PerkinElmer Wallac 1480 WIZARD 3 User manual

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INSTRUMENT MANUAL

1480 WIZARD 3"

Gamma Counter

1480-931-20

June 2004

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Wallac 1480

WIZARD3’’

Gamma counter

For instruments with software version 3.6

PerkinElmer Life and Analytical Sciences, Wallac Oy, P.O. Box 10, FIN-20101 Turku, Finland.

Tel: 358-2-2678 111. Fax: 358-2-2678 357. Website: www.perkinelmer.com

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Warning

This equipment must be installed and used in accordance with the manufacturer's recommendations. Installation and service must be performed by personnel properly trained and authorized by PerkinElmer Life and Analytical Sciences.

Failure to follow these instructions may invalidate your warranty and/or impair the safe functioning of your equipment.

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Contents

1 Introduction ............................................................................................................. 7

1.1 Introduction to PerkinElmer Life and Analytical Sciences ..................................................................7

1.2 Introduction to 1480 WIZARD ............................................................................................................7

1.3 Introduction to this manual...................................................................................................................7

CPM operation of WIZARD 3"............................................................................. 9

RiaCalc WIZ CPM operation of WIZARD 3"................................................... 11

MultiCalc operation of WIZARD 3"................................................................... 13

2 WIZARD controls.................................................................................................. 17

2.1 Introduction ........................................................................................................................................17

2.2 Keyboard ............................................................................................................................................17

2.3 Display ...............................................................................................................................................18

2.4 Live display........................................................................................................................................19

2.5 Racks..................................................................................................................................................21

2.6 Principle of the ID system..................................................................................................................21

2.7 Fitting ID labels..................................................................................................................................22

2.8 Loading racks the right way round.....................................................................................................23

2.9 Instruction labels ................................................................................................................................25

2.10 Numerical labels.................................................................................................................................25

2.11 Barcode errors ....................................................................................................................................26

2.12 Help....................................................................................................................................................26

2.13 Short-cut keys.....................................................................................................................................26

3 CPM operation....................................................................................................... 29

3.1 Introduction ........................................................................................................................................29

3.2 Start up ...............................................................................................................................................29

3.3 Normalization.....................................................................................................................................29

3.4 Protocol editing ..................................................................................................................................29

3.5 Parameters available...........................................................................................................................33

3.6 Output.................................................................................................................................................33

3.7 Leaving the editor...............................................................................................................................34

3.8 Running an assay................................................................................................................................34

3.9 The FILES function............................................................................................................................35

4 Operation with internal RiaCalc WIZ................................................................. 39

4.1 Introduction ........................................................................................................................................39

4.2 Start up ...............................................................................................................................................39

4.3 Normalization.....................................................................................................................................39

4.4 Protocol editing ..................................................................................................................................39

4.5 Parameters available...........................................................................................................................42

4.6 Output editing.....................................................................................................................................48

4.7 Leaving the editor...............................................................................................................................53

4.8 Running an assay................................................................................................................................53

4.9 The FILES function............................................................................................................................54

4.10 Selectable outputs...............................................................................................................................59

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Contents

5 Operation with external MultiCalc...................................................................... 63

5.1 Introduction ........................................................................................................................................63

5.2 Start up ...............................................................................................................................................63

5.3 MultiCalc protocol .............................................................................................................................64

5.4 Editing a MultiCalc protocol..............................................................................................................65

5.5 Running an assay................................................................................................................................68

5.6 The FILES function............................................................................................................................69

5.7 Real time clock...................................................................................................................................70

5.8 Selectable outputs...............................................................................................................................72

6 Normalization......................................................................................................... 75

6.1 Normalization.....................................................................................................................................75

6.2 GLP test normalization.......................................................................................................................79

7 Additional WIZARD functions ............................................................................ 83

7.1 Dual label counting ............................................................................................................................83

7.2 Multiple isotope assay counting .........................................................................................................87

7.3 System mode ......................................................................................................................................99

7.4 High activity mode ...........................................................................................................................113

7.5 Power failure ....................................................................................................................................115

7.6 Routine maintenance........................................................................................................................117

7.7 Safety and radioactive materials.......................................................................................................119

8 Instrument description........................................................................................ 123

8.1 Introduction ......................................................................................................................................123

8.2 FlexiRack counting ..........................................................................................................................123

8.3 Choice of energy range.....................................................................................................................123

8.4 High efficiency detector ...................................................................................................................123

8.5 Detector shielding ............................................................................................................................123

8.6 Isotope selection...............................................................................................................................124

8.7 Multichannel analyzer ......................................................................................................................124

8.8 Multiple label counting ....................................................................................................................124

8.9 High activity samples .......................................................................................................................124

8.10 Self-contained...................................................................................................................................125

8.11 Interactive control ............................................................................................................................125

8.12 Multi-user ID system........................................................................................................................125

8.13 External communication...................................................................................................................126

8.14 Multi-technology data management software ..................................................................................126

8.15 GLP performance testing..................................................................................................................126

9 Specifications........................................................................................................ 129

9.1 General description ..........................................................................................................................129

9.2 Physical dimensions .........................................................................................................................129

9.3 Electrical requirements.....................................................................................................................129

9.4 Environmental requirements ............................................................................................................129

9.5 Sample handling...............................................................................................................................129

9.6 Detector system................................................................................................................................130

9.7 Live spectrum display ......................................................................................................................132

9.8 Hard copy.........................................................................................................................................132

9.9 Max. count rate.................................................................................................................................132

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9.10 Built-in computer .............................................................................................................................132

9.11 Keyboards ........................................................................................................................................132

9.12 CRT display .....................................................................................................................................132

9.13 Operating system:.............................................................................................................................132

9.14 Multiuser Capability.........................................................................................................................132

9.15 Multitasking .....................................................................................................................................132

9.16 Helps ................................................................................................................................................132

9.17 Positive cassette information............................................................................................................133

9.18 Two simultaneous counting regions.................................................................................................133

9.19 Decay correction ..............................................................................................................................133

9.20 Contamination guards ......................................................................................................................133

9.21 Automatic power failure recovery....................................................................................................133

9.22 Date and time ...................................................................................................................................133

9.23 Connections......................................................................................................................................133

9.24 Programmable computer I/O............................................................................................................133

9.25 Datalogger ........................................................................................................................................133

9.26 Remote instrument control ...............................................................................................................133

9.27 Electrical safety requirements ..........................................................................................................133

9.28 RiaCalc WIZ, standard features .......................................................................................................134

9.29 GLP performance testing..................................................................................................................135

10 Calculation methods ............................................................................................ 139

10.1 Background normalization ...............................................................................................................139

10.2 Isotope normalization and GLP TEST measurement .......................................................................143

10.3 RIA/IRMA/RATIO assay counting..................................................................................................164

10.4 Multiple isotope assays (MIA) .........................................................................................................177

10.5 Appendix: Some basic mathematical formulas ................................................................................183

11 Installation............................................................................................................ 189

11.1 Installation procedure.......................................................................................................................189

11.2 Environment.....................................................................................................................................189

11.3 Electric power ..................................................................................................................................189

11.4 Unpacking ........................................................................................................................................189

11.5 Checking the mains voltage setting..................................................................................................192

11.6 Installing the lead shielding..............................................................................................................193

11.7 Installing the detector .......................................................................................................................194

11.8 Connecting up the counter and peripherals ......................................................................................198

11.9 Switching on WIZARD....................................................................................................................199

11.10 Installing MultiCalc..........................................................................................................................199

11.11 Functional check ..............................................................................................................................200

12 Declaration of Conformity for CE-marking

13 Index ...................................................................................................................... 207

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Trademarks

MultiCalc and WIZARD are registered trademarks and RiaCalc a trademark of PerkinElmer, Inc.

IBM and PC-DOS are registered trademarks of International Business Machines

MS-DOS, Microsoft and Excel are registered trademarks of Microsoft Corporation

DeskJet and LaserJet are trademarks of Hewlett-Packard.

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1 Introduction

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1 Introduction

1.1 Introduction to PerkinElmer Life and Analytical Sciences

PerkinElmer Life and Analytical Sciences is the world's leading manufacturer of automatic gamma counters. With the Wallac brand name it has been a pioneer in this field for many years and has a well-founded reputation for technological innovation and excellence in quality both in products and service.

1.2 Introduction to 1480 WIZARD

Wallac 1480 WIZARD 3" is an automatic gamma counter designed for counting high energy samples (up to 2000 keV). Your WIZARD can take 270 (28mm) or 1000 (13mm) samples at a time and any mixture of the two sizes of rack, since it automatically determines what the rack size is before a rack is moved into the counting position. WIZARD has its own built-in display and keyboard for full communication with its users. WIZARD uses a builtin 30 MByte hard disk.WIZARD has two energy ranges: a normal range 15 - 1000 keV and an extended range 15

- 2000 keV for higher energy isotopes.

WIZARD can operate as an automatic standalone CPM counter, or it can be used to do extensive data evaluation with its own internal RiaCalc WIZ program or it can be linked up to an external PC and use the power of the MultiCalc program.

1.3 Introduction to this manual

1.3.1 Step-by-step instruction sheets

There are three sheets following this page which give a brief step-by-step outline of how to use WIZARD in each of the three operating modes:

1 WIZARD is used as a stand-alone CPM counter. Counting parameters are set in the counter. CPM results are sent to the built-in display and a printer.

2 WIZARD is run with RiaCalc WIZ. All counting protocols are set in the counter itself. Results are sent to the built-in display and a printer.

3 WIZARD is connected to a personal computer running MultiCalc software. All counting protocols are set in the personal computer. Results are automatically returned to the PC for final evaluation and output. More information about MultiCalc will be found in the MultiCalc User Manuals.

You should find enough information on the appropriate one of these sheets for normal operation in any of these modes.

1.3.2 WIZARD controls

If you are a first time user of WIZARD you should read Part 2 "WIZARD controls" before starting to operate the counter. This part explains the basic techniques involved in using the built-in display and keyboard(s). It also explains the barcode ID system.

Once you are familiar with the techniques of using WIZARD you can proceed to actually operate it.

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1 Introduction

1.3.3 Using WIZARD to get results

WIZARD counts samples and if necessary evaluates the results following instructions given in the form of protocols (lists of parameters). In normal operation these protocols are already set-up so all you need to do is follow the instructions on one of the step-b y-step sheets. If, however, it is your responsibility to create or edit protocols then you will find t he information you need to help you in the appropriate one of the parts described below:

CPM counting requires only three parameters to be set as described in Part 3 CPM operation. RiaCalc WIZ operation is described in Part 4. Each feature of RiaCalc WIZ is described and guidelines are

given to help you use these features to achieve the results you want. If you are going to be working with MultiCalc runningonanexternalPCthenturntoPart 5. This describes

those things you need to know in order to use WIZARD and MultiCalc together. The use of MultiCalc is described in a separate User Manual which comes with the software.

Part 6 tells about Normalization. This is an operation which has to be done before WIZARD is used to count samples with a particular isotope and energy range. When it has been done once, it should be repeated occasionally e.g. after half a year. It also tells about how to do performance testing with GLP test normalization.

Part 7 describes a number of functions which are available in addition to the three main ways of using WIZARD described in parts 3, 4 and 5. These functions are Dual label, Multi-isotope assay (MIA), System functions,

High activity mode, P ower failure recovery and Routine maintenance. plus additional Safety information. Part 8 of this manual gives you a description of how WIZARD work s. You do not need this information for

normal operation but it will help you to have confidence in your results when you know how WIZARD has been designed to give you the very best.

Detailed specifications are described in Part 9 giving numerical values for e.g. efficiency, background etc. In Part 10 there is also a description of the calculation methods used in WIZARD. Part 11 contains t he information you need when installing WIZARD for the first time. Normally this will be

done by a service engineer so you will not need this information. Part 12 contains the Declaration of conformity for CE-marking to show the quality standards followed in the

making of WIZARD. Part 13 contains the alphabetical index to this manual.

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CPM operation of WIZARD®3’’

1 Fix ID clips to racks

ID labels (barcodes) are stuck to an ID clip which fits onto a rack to tell WIZARD the function of the rack. A counting protocol is a set of three parameters time, max. counts limit and isotope, which control counting. Rack numberis optional and allows each rack to have its own number. Normalization ensures that the gain of the detector is optimum for each isotope and energy range. Background ensures that the effect of the background is removed from the measured counts. Test initiates a GLP performance test normalization. Isotope number shows the isotope to be used in normalization. Stop tells WIZARD that no more racks are to be counted.

2 Load racks onto WIZARD

Make sure that tube holders are positioned correctly and that racks are loaded the correct way round with

the ID clips facing away from you as shown in the figure. Start by loading the right-hand side of the conveyor.

akesure tube holders are the right way round

Position 1

ix ID

clip here

rotocol, isotope or energy

range number

ack number or NORM, TEST, BKG or

STOP instruction

ID clip faces forward when loading racks

osition 1

3PressSTARTtocount

Live display during counting is obtained by selecting from the main menu on the WIZARD built-in display "Operate" and then "Show cpm results". Available live displays are: counting parameters, Counts, CPM, CPS and Spectrum.

ive display available

during counting

TART

4 Results are printed out

The built-in program allows counting and normalization protocols to be created and edited. The built-in display and keyboard is used. Results are sent to the display and a printer.

For Systemsetting info. e.g. detector deactivation or clock setting select SYSTEM in the main menu

The printer connected to WIZARD port 1 is used for printing corrected CPM results directly from WIZARD.

A disk can be used for transferring results from WIZARD

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RiaCalc WIZ operation of WIZARD®3’’

1 Fix ID clips to racks

2 Load racks onto WIZARD

Make sure that tube holders are positioned correctly and that racks are loaded the correct way round with

the ID clips facing away from you as shown in the figure. Start by loading the right-hand side of the conveyor.

3PressSTARTtocount

akesure tube holdersare the right way rou nd

Position 1

ix ID

clip here

rotocol, isotope or energy

range number

ack number or NORM, TEST, BKG or

STOP instruction

ID clip faces forward when loading racks

osition 1

ive display available

during counting

4 Results are printed out

The built-in program allows counting and normalization protocols to be created and edited. The built-in display and keyboard is used. Results are sent to the display and a printer.

TART

For System setting info. e.g. detector deactivation or clock setting select SYSTEM in the main menu

The

rinter connected to WIZARD port 1

is used for printing corrected CPM results directly from WIZARD

A disk can be used for transferring results from WIZARD

External keyboard in built-in drawer for extended protcol editing

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MultiCalc®operation of WIZARD®3’’

1 Fix ID clips to racks

ID labels (barcodes) are stuck to an ID clip which fits onto a rack to tell WIZARD the function of the rack. A counting protocol is a set of three parameters time, max. counts limit and isotope, which control counting. Rack numberis optional and allows each rack to have its own number. Normalization ensures that t he counting efficiency of each detector is the same. Background ensures that the effect of the background is removed from the measured counts.

Test initiates a GLP performance test normalization. Isotope number shows the isotope to be used in

normalization. Stop tells WIZARD that no more racks are to be counted.

2 Load racks onto WIZARD

Make sure racks are loaded the correct way round with the ID clips facing away from you as shown in the figure. Start by loading the right-hand side of the conveyor.

akesure tubeholdersare the rightway round

Position 1

ix ID

clip here

rotocol, isotope or energy

range number

ack number or NORM, TEST, BKG or

STOP instruction

ID clip faces forward when loading racks

osition 1

3 Press F1 (COUNTER) select 1480 and press Enter

ive display available

during counting

TART

4ResultsgotoMultiCalc

MultiCalc is a very versatile data handling program which runs on an external computer. You can use it to make counting protocols. These will be transferred to WIZARD to control counting. Results are returned to MultiCalc for evaluation. They can also be sent, via MultiCalc, to a local area network (LAN) or mainframe

Output to external computer e.g. a mainframe. Connection is from the mainframe to the PC

Local area network connection

LAN

(LAN) using the PC

Results go to the PC connected to port

2. Printout goes to the printer connected to the PC

For Systemsetting info. e.g. detector de-activation or clock setting select SYSTEM in the main menu

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

2 WIZARD

controls

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2 WIZARD controls

2.1 Introduction

This chapter describes features you need to use to control WIZARD. These features are: the one or two keyboards, the display, the ID system and the HELP function. When you understand how to use these features then you can proceed to the following chapters to see how to use WIZARD to get the results you want.

2.2 Keyboard

WIZARD can have two keyboards, a simple membrane type on the front of the instrument (see the figure below) and a second, a complete PC keyboard, which i s in a separate compartment. The instrument is normally operated using the small keyboard whereas the larger keyboard is used for extended protocol editing because the small keyboard does not include letters and certain other characters which are needed in some editing operations.

The keyboards are connected in parallel with each other and each key on the built-in keyboard has its equivalent on the external one. The keys on the external keyboard which correspond to those on the built-in keyboard are as follows:

Built-in keyboard External keyboard START F3 STOP F4 STAT F5 HELP F1 EXIT ESC C (Clear) Backspace E Enter

Use the EXIT (ESC) to escape from the operating level the program is on and go back to the previous one. If you are using a PC running MultiCalc all commands involving MultiCalc are given via the PC keyboard.

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2.3 Display

2.3.1 Main menu display

When you start to work with the instrument you will see on the built-in display something like:

==1480 Main Menu=====================

OPERATE PROTOCOL FILES SYSTEM

==Submenu===========================

Show cpm results

Show evaluationresults

Operate conveyor

==Cpm==============================

PressSTARTto measure

2.3.2 Function selection

The line in capital letters shows four major functions of which the first, OPERATE, is currently selected. The following lines show the commands available as part of that function. Each function has its own set of commands. You can select the function with the left and right arrow keys. The individual command is selected with the up and down arrow keys. To give the selected command to WI ZARD press the E key

2.3.3 Enabled and disabled functions

On the example screen shown above the first two command lines are in subdued colour. This indicates that the counter is not measuring and hence these functions are not enabled. The third item o n the menu "Operate conveyor" is however available.

2.3.4 Operating the conveyor

If you select this and press E the display changes to:

--Operate conveyor-------------------------------------

Move racks to output line

Move racks to input line Move both lines forwards Move both lines backwards Move all lines clockwise Rotate all lines counterclockwise Clear conveyor

--Press-START ---------------------------------------

By selecting from this menu and pressing E you can control the movement of racks on the conveyor. To returnto the main display, press EXIT.

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2.3.5 Selecting the current mode

WIZARD can be used in one of three modes: CPM, MultiCalc or RiaCalc WIZ. The current mode is shown on the lower part of the main menu display. The example on the previous pages shows that the Cpm mode is selected.

2.3.6 Status line

At the bottom of the screen there is a status line. This shows what is being measured. In the display example above the status line is "Press START to measure". The status line can have the following texts:

Press START to measure Measuring an assay Measuring background Normalizing Clearing the conveyor Seeking assay GLP test

2.4 Live display

When samples are being counted the display can show actual counting values, either collected counts, collected counts per minute (CPM values) or a complete isotope spectrum. The word "Live" indicates that the display is working in real time; values are updated at the same pace as counts are accumulated.

2.4.1 How to use the Live display

In order that the Live display works, the counting must be actually happening. Select the OPER ATE menu. The status line must show "Measuring assay" or a corresponding text that indicates

that counting is active. Select "Show cpm results" and press the "E" key. Counting parameters are shown (see the figure below).

2.4.2 Display modes.

There are five display modes (Note: there are two counting windows in dual channelcounting):

--Show cpm results------------------------------------Measuring now, elapsed time is = 19 s.

--Counting parameters---------------------------------Measurement Assay Protocol 11 PROT03 Label I-125 Preset time 60 Counts limit 9999999 Batch number 2

--Change data with

←→keys----------------------

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Counting parameters (as shown above) COUNTS Accumulated counts in counting window

CPM Counts per minute in the counting window CPS Counts per second in the counting window Spectrum display

Select the appropriate one using the "right" and "left" arrow keys. Pressing the right key twice initiates the CPM display

Assay (A uto mode) Measuring now ,elapsed time is = 30 s.

Isotope Count per minute

I-125 5555.3

The example is from CPM counts; the display for COUNTS and for CPS are analogous to the CPM display.

Note: CPM values are not corrected for dead time, background, spillover or detector efficiency but are direct values of accumulated counts divided by measured time.

2.4.3 Displaying the isotope spectrum

When you press”E” the spectrum is displayed. The detector spectrum display is a graphical representation of isotope activity with respect to isotope energy. The X-axis shows the energy in keV (up to 1024 or 2048 depending on the ener gy range) and the Y-axis s howsthe accumulated counts in each individual channel. The

LIVE SPECTRUM 1 X scale = MCA

total counts in full window76352

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analyzer is initially adjusted so that each channel corresponds to 1 keV, therefore the X-axis is approximately from0to1MeV.

Scale in channels or keV - The X-axis scale can be in channels or in keV. To flip from one mode to the other, press "E" when the spectrum is displayed.

2.4.4 Spectrum functions

On the right hand side of the spectrum five functions are displayed. By using the up and down arrow keys, you can select the function you want. A function is shown to be selected by the underline mark appearing under the function name. The functions are as follows:

Scale - T his allows you to select the scale for the spectrum. The default is 1 - 1024. By pressing the le ft arrow you can change the scale to 0..512, 0..256, 0..128, or 0..64. To return to fuller scales, press the right arrow. The figure shows the 0..64 scale. See the next function "Shift" for how to move through the scale segments.

Shift - If the scale used is less t han full scale (0 - 1024 or 1 – 2048, e.g. 0 - 256), you can use the "shift" function to move the spectrum to see other parts which otherwise would not be displayed e.g. 256 - 512 etc.

Marker – If you have one of the basic scales (1 – 1024 or 1 – 2048) selected you can use this function to select a marker which you can then move with the arrow keys to mark any position in the spectrum. In this way you can find out the exact position of any peak.

WinLo and WinHi - These allow you to set an upper and lower window limit marker so that you can obtain the

counts within the window. You can also print out the spectrum if you press "Print screen" on the external keyboard provided CAPS LOCK

is not on and there is a printer connected. Spectra can also be printed via MultiCalc to the printer that is connected to the PC running MultiCalc. To do this, the following conditions must be met: If the instrument is in "MultiCalc" mode, then MultiCalc itself

must be in online mode when the Print Screen key is pressed. If the instrument is in "Cpm" or "RiaCalc WIZ" mode, then the instrument parameter "SYSTEM |Printout options | Without buffering PC" must be "YES" and MultiCalc must be receiving data from the counter.

Spectra are printed correctly onlyif in the WIZARD communication protocol the Terminal parameter is VT-52.

2.5 Racks

WIZARD takes two types of rack, one which takes 5 x 28mm samples and the other which takes 10 x 13mm samples, see the figures of the racks on page 24. You can have any mix of these racks on t he conveyor because WIZARD automatically determines the rack type and adjuststhe sample loading mechanism each time a rack is taken in to the loading position.

2.6 Principle of the ID system

WIZARD is an automatic gamma counter. This means that samples from several assays can be loaded onto the conveyor and WIZARD can be left to count them by itself. To do this it needs to be able to identify each batch of samples. The WIZARD ID system uses a plastic clip onto which one or two barcode labels are stuck. The clip is then clipped onto the rack to be identified. The clip can easily be removed and a new one fitted when required.

The barcodes give information to WIZARD aboute.g. which counting protocol is to be used for the samples, what the rack number is etc. Only the f irst rack in an assay needs to have a protocol label. Barcodes are to be found in a booklet supplied with the counter. There are two classes of barcodes, numerical ones (0 ... 99) and instruction

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2 Wizard controls

ones. The figure above shows an empty clip and then several examples of clips with labels on them. Some have two labels and some only one.

In addition there is a third area of the clip which comes on the end of the rack. This can be used for you to write your own information on. The ID label booklet includes empty labels for you to write on and stick to the clip. There are also barcodes marked P CURVE and CONTROL which are not used in this program.

Empty clip

Clip with rack number (02) and protocol number (18)

Clip with normalization instruction and isotope code number 02

Clip indicating a background measurement to be made

Clip indicating that counting is to stop after counting this rack if it contains samples. If it is empty counting will stop immediately

2.7 Fitting ID labels

The figures below show two types of rack with an empty ID clip fitted on them. There is a special recess on one side of each rack. This is where the clip fits. It will not fit to the other end of the rack. This is important because it defines which way round the rack should be on the conveyor. Make sure the ID clip is fitted properly so that it does not slip off when the rack is on the conveyor.

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Empty clip fitted onto rack

2 WIZARD controls

Empty clip fitted onto rack

2.8 Loading racks the right way round

When you load racks on the conveyor the ID labels must face away from you with the white dot on the right side as shown in the figures on the next page. The handwritten information on the label at the end of the rack is then clearly visible from the side of the conveyor.

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All racks, even those without labels, must be put the same way round with the recess for the label (and white spot)on the right hand side when you load the racks. If a rack is not the right way round a warning is displayed and counting is stopped until the rack has been turned the right way round.

Make sure the tube holders are the right way round, see the figures.

Direction of movement on the conveyor

White dot

Rack orientation when loaded onto the conveyor

Direction of movement on the conveyor

Rack orientation when loaded on the conveyor

Information written by the user on the ID label

White dot

Information written by the user on the ID label

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2 WIZARD controls

2.9 Instruction labels

Use instruction labels to select special functions; attach the label in the area marked RACK/SPECIAL. The instrument recognizes the following codes:

2.9.1 STOP

This stops counter operation after the counting of the rack with this label. An alternative is to use an empty rack as a STOP rack.

2.9.2 NORM (Normalization)

This label is used i n conjunction with the isotope code label (see 2.10.2 below) to identify a rack as a normalization rack. The isotope must be in the last position in a rack which is position 10 (or position 5 in the large volume racks). Other positions must be empty and without holders. See chapter 6 for detailed instructions.

2.9.3 BKG (Background)

A rack with this label is used to make background normalization. No sample tubes should be in the rack, see chapter 6. WIZARD has two energy ranges 15 - 1000 keV and 15 - 2000 keV. If you only use a BKG label then normalization will be done for both energy ranges. If you include an odd or even numeric label on the PROTOCOL area of the clip then only the normal or extended energy range respectively will be normalized.

2.9.4 TEST (GLP test)

This label is used for coding the GLP (Good Laboratory Practice) test normalization. The isotope must be in position 10 (the last position in a rack). Other positions must be empty and without holders. See chapter 6 for detailed instructions.

Note: Make sure you remove NORM, BKG and TEST racks after use to avoid redoing a normalization unintentionally.

2.9.5 SPECIAL

This label is used to select the High activity mode. See Chapter 7.4 for more details.

2.10 Numerical labels

2.10.1 Protocol number label

Protocols (1.. 99) are called into use by numerical barcodes, the number refers directly to protocol number. The label should be attached to the area marked PROTOCOL.

Although the ID system is used, not all racks need have ID labels on them. In automatic counting the racks are counted according to the last protocol number label until the next protocol number label or a special code is read.

2.10.2 Isotope number code label

The number of the isotope to be used in normalization must be shown by attaching an appropriate numerical label to the PROTOCOL area when the rack is being used as a normalization rack.

2.10.3 Rack number label

Use numerical labels 1 .. 99 to select the rack number, attach the label in the area marked RACK/SPECIAL. This is optional and need not be used at all.

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2.10.4 Energy range selection label

This is an odd or even numeric label, attached to the PROTOCOL area in conjunction with the BKG label as described above in section 2.9.3.

2.11 Barcode errors

If the ID system fails to read the barcode successfully the rack is handled as if there was no barcode. If the code is read successfully but that particular protocol does not exist the message "Protocol not found" appears on the display and printer and the counter stops.

If there are two numerical codes on the same ID clip, the one in the RACK/SPECIAL area is taken as the rack number and the one in the PROTOCOL area as the protocol number.

2.12 Help

The WIZARD software includes an extensive context sensitive Help function. If at any point while operating WIZARD, you are not sure what to do, or what a particular function is, just press the HELP key on the built-in keyboard (or F1 on the external keyboard). An explanation of the currently selected feature will appear on the display.

When you have read the help text you can go back to the function about which you requested help by pressing the EXIT key.

2.13 Short-cut keys

Two short-cut keys Alt-I and Alt-E are available on the main menus level: Alt-I (the I meaning "internal") sets the mode to "RiaCalc WIZ" and the SYSTEM parameter Printout

selections/Write results to file" to "Yes". Alt-E (the E meaning "external") sets the mode to "MultiCalc" and SYSTEM parameter Printout selections/Write

results to file" to "No". These short-cut keys are available only when S YSTEM parameters can be edited (that is, when measurement is

not occurring).

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3.1 Introduction

The instructions here describe the routine operation of WIZARD when it is running in CPM mode.

3.2 Start up

1. Switch on the printer; this should already be connected to the counter.

2. You can put the data disk named 1480 Datadisk into the WIZARD disk drive. The name label should be facing upwards.

3. Switch on WIZARD. After about 3 min. the display will show:

==1480 Main Menu=====================

OPERATE PROTOCOL FILES SYSTEM

==Submenu===========================

Show cpm results

Show evaluationresults

Operate conveyor

==Cpm==============================

PressSTARTto measure

4. Check the time and date by selecting SYSTEM mode and DATE. If it is not correct then give the correct value (See "System | Time & Date setting"). Return to the main display by pressing EXIT.

5. Make sure that the counter is in the right mode i.e. that the text CPM appears near the bottom of the screen as shown in the example above. If it does not then select "System" and then the operation mode. The mode must be CPM.

3.3 Normalization

Make sure that WIZARD has been normalized for the isotope you are going to use in your measurement. See chapter 6 "Normalization".

3.4 Protocol editing

3.4.1 What is a protocol?

A CPM protocol contains three parameters for controlling counting conditions. Before counting can be done, a protocol must exist. Each protocol must have a name and an ID number. This

number allows the protocol to be called into use by means of a barcode label. To access protocol file handling you must first select the PROTOCOL option on the display.

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3.4.2 Protocol file handling

There are several commands available to use with a protocol, the most common ones are CREATE (making a new protocol) and EDIT (change the contents of the existing protocol).The list that appears after you select PROTOCOL is as follows.

==1480 Main Menu===================== OPERATE ==Submenu===========================

==Cpm=====================More

PROTOCOL FILES SYSTEM

Create

Edit

Copy Rename Delete Recover Print Load

↓ ===

PressSTARTto measure

Create

Select CREATE and press E. Give the protocol name. There is a default name of the form "Protnn" where "nn" is the protocol number. If you want to change this name you must use the external keyboard unless you only use numbers for the name. A maximum of eight characters can be given. Select an unused ID number (between 1 to

99). Press E. The protocol parameters can be set in a similar way to those in editing.

Edit

Select Edit and press E. Select the protocol by name from the list of protocols available and press "E".

--Edit protocol---------------------------More ↑-----05 BF_BLAN RIA 06 FOLATE RIA 07 FORMAT RATIO 08 LEARN RIA 09 PROT01 RIA 10 PROT02 RIA

11 PROT03 RATIO

12 READ_ME RATIO 13 PROT00 RIA 14 TEST_1 RIA 15 TEST_3 RIA 16 TSH RIA

--Id order, change with

←→-----------More ↓ ------

You may now change the parameters. You can move up and down in the parameter list by means of the UP and DOWN arrow keys. See section 3.5 for details of parameters available in a protocol.

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Note: In the example above the protocol types include both RIA and RATIO. It is not possible to create RIA protocols in CPM mode (protocols are automatically labelled RATIO) but if such a protocol has been created in another mode then it will still appear on the list. You can select any protocol from the list for editing in CPM mode but only the three p arameters shown in the example below will appear.

--Edit protocol------------------------------------------11 PROT03 RATIO

Counting time 60

Max. counts limit 9999999 Labels I-125

___________________ ___ ___ ___ ___ ____ __ __

Copy

An existing protocol can be copied so as to create another protocol with the same contents. Select Copy and press E. Select the protocol to be copied. Give the name the new protocol is to have. You may

also give an ID number. Select "Do copy" and press "E".

Rename

Select this option to give a new name to a protocol. Select Rename and press E. Select the protocol to be renamed from the list of protocols. Give it a new name and/or ID number. Select "Do rename" and press "E".

Delete

Select Delete and press E. Select the first protocol to be deleted. Then select "Do delete" and press "E". The protocol and any associated data willthen be deleted.

Recover

Deleted protocols can in most cases be recovered by selecting this option. They are saved in the protocol index area. There is room on the system disk for 99 deleted and active protocols. The larger the number of active protocols the fewer the number of deleted protocols that can be saved. If there is no room, the oldest deleted protocol will be permanently removed to make room for a newly created one. No protocol with the same name should have been deleted later than the one you want to recover.

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With this function you can print the contents of a single protocol or the protocol index according to the selection you make.

Isotope names are included in t he protocol printout. For dual label protocols the code numbers for both isotopes are printed separately, e.g. 1; I-125 + 2; Co-57.

Load

You can load a single protocol or all protocols from an external microfloppy disk. Such protocols will have been saved there using the SAVE function (see below). Note: if you load a new protocol when a previous one with the same name or ID number exists in the instrument, you have the choice of either renaming the protocol to be loaded or deleting the previous protocol.

Normal Load will give you: 1 Protocol files

2 Controls files 3 Trends files

Extended Load will in addition give you: 4 Standard curve files

5 Data files 6 Results files

Note: if you load a new protocol when a previous one with the same name or ID number exists in the instrument, you have the choice of either renaming the protocol to be loaded or deleting the previous protocol.

Save

This function allows you to transfer files from WIZARD to an external microfloppy disk for storage. Subsequently these files can either be loaded back into WIZARD.

Purge

This function permanently erases deleted files from the instrument hard disk. You cannot recover them at all.

Password

When a protocol is created, you can give a two-character password. All characters are allowed in the password and the password is case sensitive. If you do not give a password at this point, then the protocol can be edited, renamed, overwritten when a new protocol with the same name or ID number is loaded and deleted without giving any password. Otherwise the password is needed to do these operations. A protocol can always be copied, restored, saved and purged without giving the password. With rename it is possible to change or remove the password.

Passwords are retained even if power is turned off. The password feature can be disabled by removing the file C:\PASSWORD from the instrument hard disk. To enable it again, type ECHO aa > C:\PASSWORD at the DOS prompt to recreate a password file and boot the instrument. (You can enter DOS by installing the installation disk to the disk drive and by restarting the instrument.)

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3.5 Parameters available

3.5.1 Counting time

Give the counting time in seconds. All samples are counted for this time. The d efa ult value is 60 and the maximum value is 65000 seconds.

3.5.2 Max. counts limit

If you want to terminate counting on the basis of the number of counts accumulated, enter the counts value for this parameter. Provided this number of counts is reached in each detector before the counting time expires, this parameter will terminate counting. If however the counting time expires first it will terminate counting even though the max. counts limit has not been reached. The maximum and default value for this parameter is

99999999.

3.5.3 Labels

If the isotope you are using is I-125 just press the down arrow key. If you use another isotope change the isotope with the "left" arrow key or by pressing E. The isotope list is shown in chapter 8.2 Specifications.

To change the contents of this isotope list go to the SYSTEM mode and the Isotopes parameter, see charpter 7.3 in this manual.

If you want to do dual label counting select the second isotope with the right arrow key and press E or press E to get the label selection menu.

3.6 Output

3.6.1 Default output

In CPM mode there is a default format for output and this cannot be changed. The items displayed and printed are: Sample position, rack number, batch number, counting time, counts, CPM and CPM error. Note: The registered counts in a counting region (window) are printed out as "COUNTS". These values do not

include background subtraction or any other correction. The following corrections are applied to get the final CPM value:

- background correction (if background normalization done)

- dead time correction

- decay correction (if selected)

- spillover correction (dual label assays)

The CPM values shown in the live display are uncorrected.

In addition there are:

3.6.2 Start time

The exact time (to the nearest 1/10 second) when measurement of each sample was started can be included in CPM printouts.

If this starting time output has been selected, a field named "CLOCK" appears at the right end of the CPM printout.

The measurement start time field can be enabled and disabled in the following way.

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Set the SYSTEM parameter Diagnostic output | Print meas. start time" to "Yes" to enable the field and to "No" to disable it.

3.6.3 Run ID

Each time a batch of samples is run it is given a run ID. This will be printed at the beginning of the CPM results. The run ID is specific for each protocol. This enables you to distinguish multiple runs of the samples with the same protocol ID.

3.6.4 Dead time factor

The Dead time factor can be included in the CPM printout. To do this, set the SYSTEM parameter "Diagnostic output | Print dead time factor" to "Yes". Dead time factor is explai ned in 8.3.1.3.

3.6.5 Bad spectrum results

Note: the output values for counts and CPM are 0 if the spectrum is bad e.g. if the coincidence peak is missing or too small for I-125.

3.7 Leaving the editor

Press "EXIT" to leave the editor A choice of three possibilities is shown:

Save changes and exit. Saves the parameter setting on file and leaves the editor. Quit and ignore changes. Leaves the editor without saving the changes. Edit. You return to the editor to do further editing.

3.8 Running an assay

Make sure that the first cassette has its correct protocol selection ID and that the appropriate protocol is stored in the instrument. For more information about the " ID system" see section 2.1.5. Put a "STOP" ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically.

Load racks, starting with the right-hand conveyor lane. Start counting by pressing the START key. The instrument will now count all the loaded samples and the WIZARD CPM software will print out the CPM

results. During counting you can see the live results on the live display as described in section 2.1.4 by selecting the menu

item "Show CPM results" to see the output from WIZARD on the display. The counting will stop automatically when a "STOP" rack is found. You can also press the STOP key on the

WIZARD keyboard. In that case the following text will appear:

Continue End assay, continue End assay, clear conveyor

Depending on whether you want the next assay to be counted or all counting to stop, select "End assay, continue" or "End assay, clear conveyor" respectively. To override the stop instruction and continue counting select "Continue".

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3.9 The FILES function

Most FILES functions are not available in CPM mode, (they are in grey), they are only available in RiaCalc WIZ, however the following three are:

3.9.1 Spectra

Store spectra - this is for information only. It tells if spectra can be saved or not and if they are to be sent to an

output device or not. The actual settings can be made in SYSTEM | Operation mode | Store assay spectra. See there for more details.

Operation - this allows you to handle the files of spectrum data. The options are: Delete - delete the spectrum file data Save to disk - save the spectrum information on the program disk in WIZARD Send to PC - send the spectrum information to a PC that is connected to WIZARD

3.9.2 GLP data

This allows you to handle the GLP data obtained in GLP test normalization. There are four options, some of which lead to other options:

View - allows you to view the GLP data Delete - delete GLP data Criteria - allows you to select the warning limits for different types of GLP data. See Item below Isotope - you can select the isotope type from those for which a GLP test normalization has been enabled Item - many items appear from which you can select the one which you want the GLP data: PEAK, BGRD,

RESOL, EFFICIENCY, COVERAGE, CHI-PROB, WIN-CPM, TOTAL CPM

Print criteria - select the isotope for which you want the criteria to be printed out.

3.9.3 Waste log file

A waste log file that contains the total CPS and DPS values of all measured assays, isotope normalizations and GLP TEST measurements can be printed or stored on a datalogger disk.

The file can also be deleted or sent via the PC port to an external PC. The waste log can contain approximately 700 entries, after this the older half of the entries is deleted and the log

starts growing again. For isotope normalization and GLP test, TOTAL CPS is the average corrected sample activity in the isotope

counting window in all detectors used. It is corrected for dead time, background activity and isotope decay. TOTAL DPS is the average corrected sample activity in the open window divided by Efficiency% which is a SYSTEM parameter that appears after you se lect Isotope and then the isotope name.

For assay measurement TOTAL CPS is the sum of all printed corrected CPM values of measured tubes in the assay converted to CPS. The DPS value is obtained by dividing the printed corrected CPM (converted to CPS) by the actual coverage o f the isotope counting window a nd by the parameter Efficiency% refered to above. TOTAL DPS is thus the sum of all DPS values of measured tubes in the assay.

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4.1 Introduction

The instructions here describe the routine operation of WIZARD when it is running the internal RiaCalc WIZ software.

4.2 Start up

1. Switch on the printer; this should have already been connected to the counter.

2. You can put the data disk named 1480 Datadisk into the WIZARD disk drive. The name label should be facing upwards.

3. Switch on WIZARD. After about 3 min. the display will show:

==1480 Main Menu=====================

OPERATE PROTOCOL FILES SYSTEM

==Submenu===========================

Show cpm results

Show evaluationresults

Operate conveyor

==RiaCalc WIZ=======================

PressSTARTto measure

4. Check the time and date by selecting SYSTEM mode and DATE. If it is not correct then give the correct value

(See "System | Time & Date setting"). Return to the main display by pressing EXIT.

5. Make sure that the counter is in the right mode i.e. that the text RiaCalc WIZ appears near the bottom of the

screen as shown in the example above. If it does not then select SYSTEM and then Operation mode. The evaluation must be "RiaCalc WIZ".

4.3 Normalization

Make sure that WIZARD has been normalized for the isotope you are going to use in your measurement. See chapter 6 "Normalization".

4.4 Protocol editing

4.4.1 What is a protocol?

The conditions controlling samples e.g. counting time, curve fitting method etc. are stored in a protocol. Before counting can be done, a protocol must exist. Each protocol must have a name and an ID number. This

number allows the protocol to be called into use by means of a barcode label. To access protocol file handling you must first select the PROTOCOL option on the display.

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4.4.2 Protocol operations

There are several commands available to use with a protocol, the most common ones are CREATE (m aking a new protocol) and EDIT (change the contents of an existing protocol). T he complete list that appears after you select PROTOCOL is as follows.

==1480 Main Menu==================== OPERATE ==Submenu==========================

==RiaCalc WIZ============= More

PROTOCOL FILES SYSTEM

Create

Edit

Copy Rename Delete Recover Print Load

↓ ==

PressSTARTto measure

Create

99). Give the assay type, RIA or IRMA. If you just want to get CPM results or you do screening, select RATIO. Press E. The protocol parameters can be set in a similar way to those in editing, see below.

Edit

Select Edit and press E (Enter). Select the protocol by name from the list of protocols available and press "E".

--Edit protocol---------------------------More ↑-----05 BF_BLAN RIA 06 FOLATE RIA 07 FORMAT RATIO 08 LEARN RIA 09 PROT01 RIA 10 PROT02 RIA

11 PROT03 RATIO

12 READ_ME RATIO 13 PROT00 RIA 14 TEST_1 RIA 15 TEST_3 RIA 16 TSH RIA

--Id order, change with

←→-----------More ↓ ------

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You may now change the parameters. You can move up and down in the parameter list by means of the UP and DOWN arrow keys. See section 4.5 for details of parameters available in a protocol.

--Edit protocol------------------------------------------11 PROT03 RIA

Dual evaluation NO

Counting time 60 Max. counts limit 9999999 Labels I-125 X-axis (conc) L OG Y-axis (resp) B/B0 Fitting algorithm AuSpline Std. outlier reject NO Controls NO Display

--Choice, use

←→ or ENTER--- More ↓ -----------

Copy

also give an ID number. Select "Do copy" and press "E".

Rename

Select this option to give a new name to a protocol. To do this first select Rename and press E. Then select the protocol to be renamed from the list of protocols. Give it a new name and/or ID number. Select "Do rename" and press "E".

Delete

Select Delete and press E. Then select the first protocol to be deleted. Next select "Do delete" and press "E". The protocol and any associated data willthen be deleted.

Recover

Deleted protocols can in most cases be recovered by selecting this option. They are saved in the protocol index area. There is room on the system disk for 99 deleted and active protocols. The larger the number of active protocols the fewer deleted protocols can be saved. If there is no room, the oldest deleted protocol will be permanently removed to make room for a newly created one. No protocol with the same name should have been deleted later than the one you want to recover.

With this function you can print the contents of a single protocol or the protocol index according to the selection you make.

Isotope names are included in t he protocol printout. For dual label protocols the code numbers for both isotopes are printed separately, e.g. 1; I-125 + 2; Co-57.

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Load

You can load a single protocol or all protocols from an e xternal microfloppy disk. Such protocols will have either been saved there using the Save function (see below) or will be from MultiCalc. In addition to the protocols themselves any associated data will also be loaded according to what you select as follows:

Normal Load will give you: 1 Protocol files

2 Controls files 3 Trends files

Extended Load will in addition give you: 4 Standard curve files

5 Data files 6 Results files

Save

This function allows you to transfer files from WIZARD to an external microfloppy disk for storage. Subsequently these files can either be loaded back into WIZARD or into MultiCalc. As with Load there are two options Save or Extended Save. In the former case only protocols, controls and trends will be saved. In the latter case standard curves, data, precision profiles, input files and results files will be saved also.

Purge

This function permanently erases deleted files from the instrument hard disk. You cannot recover them at all.

Password

When a protocol is created, you can give a two-character password. All characters are allowed in the password and the password is case sensitive. If you do not give a password at this point, then the protocol can be edited, renamed, overwritten when a new protocol with the same name or ID number is loaded, and deleted without giving any password. Otherwise the password is needed to do these operations. A protocol can always be copied, restored, saved and purged without giving the password. With rename it is possible to change or remove the password.

4.5 Parameters available

4.5.1 Dual evaluation

If you do normal single isotope RIA/IRMA select NO. If you would like to measure samples labelled with two isotopes in the same vial select YES. See "Dual label counting" for more details.

4.5.2 Counting time

Give the counting ti me in seconds. All samples are counted for this time.

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4.5.3 Max. counts limit

99999999.

4.5.4 Labels

The default selection is single label I-125. If you want a different isotope, press E, then specify if you want d ual label or not, then select the isotope(s) from the isotope list (see the table in chapter 9 Specifications).

To change the contents of this isotope list go to the SYSTEM mode and the Isotopes parameter, see charpter 7.3 in this manual.

For dual label counting you must also select the second isotope with the right arrow key or press E to get the label selection menu.

4.5.5 X-axis (conc)

Choose the scale o f the X-axis of the standard curve. The choices are linear, logarithmic or linear/logarithmic, i.e. linear fitting and logarithmic plotting. Logarithmic scales are commonly used.

4.5.6 Y-axis (resp.)

Choose the scale of the Y-axis from the list:

CPM counts per minute B(bound) CPM - B(blank) B/B0 (CPM-BLANK)/(BLANK-REFER) LOGIT ln (r/(1-r)), r=B/B0 LOG_B lg (CPM-BLANK) B/T CPM-BLANK)/(TOTAL-BLANK) B0/B inverse of B/B0 T/B inverse of B/T B/F r/(TOTAL-r), r=(CPM-BLANK) F/B inverse of B/F PROGR customized response PROG/LOG PROG with logarithmic coordinates

In this list B(bound) is the response value, B0 is the reference sample's response value, with a BLANK (BLANK = NSB = Non Specific Binding). The commonly used responses are B/B0 and LOGIT. T is the total sample response value. In RIA it is presumed that T >B0 >X >BLANK where X is the response of any unknown or control value.

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4.5.7 Fitting algorithm

Choose the fitting algorithm for the standard curve from the list:

LinInt linear interpolation LinUwReg linear unweighted regression ParUwReg parabolic unweighted regr CubUwReg cubic unweighted regression LinWgReg linear weighted regression ParWgReg parabolic weighted regr CubWgReg cubic weighted regression InSpline interpolated spline AuSpline auto. smoothed spline SmSpline manually smoothed spline

4.5.8 Std outlier reject

Select NO if there is to be no check for outliers but if you select CONDITION then two more parameter lines appear:

Diff. from mean 20(%) and 400 Diff. from curve 10(%)

The first specifies that an outlier will be rejected if its percentage difference and the absolute difference from the mean of the replicates equals or exceeds the value given here. The second uses a standard curve as a reference instead of the mean. You can also speci fy whether the larger or smaller of two replicates is to be the one that is rejected.

4.5.9 Curve edit halt

Choices are:

YES halt for curve edit NO do not halt for curve edit REF.CURVE evaluate using reference curve

HALT = YES causes halt, meaning that the evaluation is suspended after the standard curve until accepted by the user. The third choice does not cause halt but uses the stored reference curve for evaluation.

4.5.10 Controls

If you do not want to use control samples select NO. If you select YES then t he following parameters appear: Field For control samples normally the concentration of each control is selected, however it is possible to select a

different quantity and to specify whether it applies to individual controls, average values or averages of dilution series averages.

The next three lines allow target values to be set for the three types of controls: Low, Medium and High. followed by the additional controls Control4, Control5 and Control6. In each case you can give a target value for the control, the upper or lower 2SD limit or both plus and minus 2SD limits.

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List This parameter lets you specify a list of controls and the number of patients between controls, e.g.

<start of assay> LOW CONTROL 10 SAMPLES MEDIUM CONTROL 10 SAMPLES

You can set at most 12 controls of one kind (e.g. LOW) in the list. The list should be made long enough to allow the evaluation of assays of varying lengths. If you want the assay to end with controls you must adjust the number of patients in the list before each assay. If you edit this protocol in an external MultiCalc, you can put your own codes in the list, but they cannot be stored.

Replicates Lets you specify a replicate value for controls (1..99), but no dilution. Alternatively, you can specify that the patient replicate and dilution from the coding part be used.

4.5.11 Saved files

You can specify which types of files are required. These protocols are generated automatically during evaluation. The choices are:

Input data The data used for analysis. See Ria evaluation

from file

Standard curve Standard curve

Trend data Results for trend profiles

Control samples data Concentration results of control samples

Results data Results for output to an external computer. This

file is an ASCII file and its contents are selected on the line Result file

The default for each file type is NO (not selected).

4.5.12 Display

You can specify which results are to be displayed on the screen. Choose the ones you want.

4.5.13 Printer

This allows you to specify the results for printout. The list of items which from which selection can be made are the same as the list for the display but the actual selection for the printer may be different from that for the display.

4.5.14 Results files

This allows you to specify the contents of the Results (ASCII) file. The possibilities are the same as those for the display but the actual selection for the res ults files may be different from that for the display.

See the section entitled "Output editing" for a description of the types of output options available and how to select and edit them.

4.5.15 Coding

The program must know in which order RIA standards are loaded and what their nominal concentrations are. This information is supplied in the CODING. Other special samples such as Blank and Reference can also be specified.

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When you are creating a new protocol there is a default coding available as follows:

<Start of assay> 2 BLANK 2 REFER 2 TOTAL 2STD1=1 2STD2=2 2STD3=4 2STD4=8 2STD5=16 2STD6=32 2STD7=64 2UNKN=1

The number on the left is the replicate number and the number in the right is the nominal concentration. The replicate number preceding UNKN1 means that all unknowns have this number of replicates. In the example above all samples are duplicates.

To change a replicate number use the +/- keys to increase or decrease respectively the number of replicates. Use the numerical keys to change concentration values. Use the delete key to remove a value. If you want to edit these parameters or anyot her already created set of parameters use the cursor control keys to

select the item you want edit and press Enter. This will cause a display like the following to appear:

--Edit protocol---------------------------------------11 PROT03 RIA

--Coding-----------------------------------------------

--Edit item-------- ------------------------------------

Select what to do with the item that was selected in the coding list.. ___________________ ___ ___ ___ ___ ___ ___ __ New replicate No. 02

New item type REFER

Save replicate array Yes Cut this item Paste item after this one Make new item after this one

--Choice, use

←→or ENTER---------------------

In the example REFER was the item selected to be edited. If you want to change the coding item type select it and press E. You will then get a list of all possible item types.

Select the one you want and press EXIT twice.

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--Edit protocol --------------------------------------11 PROT03 RIA

--Coding-----------------------------------------------

--Edit item---------------------------------------------

--New item type--------------------------------------

REFER response whenconc. = 0

TOTAL total labelled antigen BLANK non specific binding POS positivedose limit NEG Negative dose limit STD Standard dose sample UNKN unknown sam ple (and dilution) REPEATremainder of samples n times

--Select and press ENT ER--------------------------

4 Operation with internal RiaCalc WIZ

The main control list will then show the new selection you have made. The recommended order for sample tubes is: blanks (NSB)

totals references (zero sample) standard sam ples unknowns.

Unknowns are either patient samples or control samples in arbitrary order, the controls must be further specified on the line called Controls.

There is also a parameter "Repeat remainder of samples n times" where n can have the values 1 to 32767 or infinite. This allows you to repeat count those samples which follow the point in the list at which this parameter is placed.

4.5.16 Options

This parameter leads to the following: Unkn.%CV flag limit A flag can be set for unknowns by typing the %CV of the concentration above which the

flag should appear. The flag is " %CV !". If you select "Not used" then the flag is disabled. Unkn. multipl. factor The value you enter for this option will be used to multiply the unknown response values.

The default value is 1. You can select "Not used" for this. Unkn. outlier limit Give a %CV value. Any unknown which has a %CV exceeding this value is not included in

the precision profile. You can select "Not used" for this.

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Trends If y ou want trends to be printed they should be selected here. The possibilities are:

Slope at ed-50 Y intercept Estimated concentration at 20% binding Estimated concentration at 50% binding Estimated concentration at 80% binding Blank over Total ratio Reference over Total ratio Blank over Reference ratio Minimum detectable concentration Parallelism factor Difference slope Histogram

The default setting for each is NO.

4.5.17 Factors

You can set the values for the factors UNIT and UNIT_B to show what the units of the results are.

4.6 Output editing

4.6.1 Output media

The protocol lines Printer, Display and Results allow you to specify which results are to be printed on the paper, or displayed on the screen or saved as an ASCII file respectively. Output selection is specific for each counting protocol. The options are the same for each output medium, o nly the medium itself (printout device, built-in display or disk) is different. In what follows only Printer selection is explained because Display and Result selections are identical to it.

Note: The registered counts in a counting region (window) are printed out as “COUNTS”. These values do not include background subtraction or any other correction. The following corrections are applied to get the final CPM value:

- background correction (if background normalization done)

- dead ti me correction

- decay correction (if selected)

- spillover correction (dual label assays) The CPM values shown in the live display are uncorrected.

4.6.2 Default or customized outputs

The program has a default set of output items (shown in the sections following) which it prints out if further instructions are not given. Therefore when you create a new protocol it is not necessary to change the printout format. However, you can choose a new type of output from a wide range of the possibilities. To alter the printout form proceed as follows:

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Select Printer in the protocol and press "E" . A menu will appear on the screen:

Printout options Printout fields Printout switches Copy settings from a template.

Each of these selections is described below.

4.6.3 Printout options

Printout options allow certain types of output to be selected. Most of these options are plots but it is also possible to have the protocol listed as well as the protocol identifier. When you select this option, a list appears with the default selection as follows:

Protocol id number YES Protocol NO Standards in table format YES Std.curve in graphical format YES Comparison curve NO Controls in table format NO Controls in graphical format NO Response error relationship NO Prec. prof. in table format NO Precision profile in graphical format NO Trends NO Histogram NO

"Protocol id number" prints the protocol name and date and "Protocol" prints the protocol contents. Results for standards can be output as curves a nd/or tables of values. Results for controls can be output as plots or

tables of values where the most recent control value is the first ite m in the table. The age of the results i ncreases as you go down the items in the table.

Trend values are output as plots. Precision profile output can be as tables or plots. A plot of the response error relationship and a histogram of results are also selectable. Select YES or NO by pressing the left or right arrow keys. Press "EXIT" when ready.

4.6.4 Printout fields

Printout fields comprises numeric values and flags. These are printed for individual samples unlike printout options which are printed for a complete assay.

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Select this option and you will see a list of the default settings:

<Left margin of paper or display> SEQ sequence or tube number PAT patient number for unknowns. CODE type in coding or contr. list TIME counting time in seconds CPM counts per minute (chn.A) CONC concentration (chn.A) %CV coefficient of variation FLAG concentration flag (chn.A)

The items are printed from left to right, a single ro w of results for each sample. If replicate samples are employed then the average results are printed, on a separate line, after individual sample values. You can delete an item from the list or add new items to the list. See section 4.10 at the end of this chapter for a descriptionof all the items available.

4.6.5 Deleting a printout field

Assuming for example that you want to delete the field FLAG from the list. Select FLAG and press "E" A menu will appear on the screen:

Change item type FLAG Cut this item Paste item after this one Make new item after this one

Note: the "paste item" line will only appear if an item has previously been deleted. Select "Cut this item" and press "E". The program will then show the new list; FLAG is not available any longer.

4.6.6 Inserting a printout field

Lets assume that you want to insert the field ETIME to the previous list, between "CPM" and "CONC". Select "CPM" and press "EDIT" A menu will appear on the screen:

Change item type CPM Cut this item Paste item after this one Make new item after this one

Select "Make ne w item after this one" and press "E" twice. A list output items will appear on the screen. See the list on the previous page.

Use the arrow keys to move through the list to the item you want. e.g. in this case "ETIME" is the sixth item on the list.

Note: You can also use the following keys on the external keyboard to speed up accessing of items: Home, End, Page Up and Page Down.

Press Exit twice.

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Notice that the part of the original list:

CPM counts per minute (chn A) CONC concentration (chn A)

has changed to a new list:

CPM counts per minute (chn A) ETIME elapsed time in decimal hours CONC concentration (chn A)

4.6.7 Pasting a cut item

The item most recently deleted with the "cut item" instruction can then be pasted back by selecting "Paste item after this one".

4.6.8 Tabulation of output fields.

Each printout field has a maximum field length, for example "time" has a length of 5 characters. It takes this space even if the number o f digits in the actual printout is less. A field is separated from the previous one by one space from the previous value. You can also define the position of the output field by giving a number called a tabulation number.

There are two approaches to tabulation: either giving an absolute position on a line in terms of the number of characters from the beginning of the line to the beginning of the field, or the number of characters between the beginning of the field and the end of the previous one. The former is called absolute tabulation and the latter relative tabulation.

The default setting is one space from the previous value. Example: There is the selection:

SEQ sequence number PAT patient number TIME counting time in seconds CONC concentration value

This is printed as follows:

SEQ PAT TIME CONC #### #### ##### ####.###

where the # mark is used here to define the field length. Default field lengths are listed in section 4.10. Assume now that you want the SEQ printout field to start from position 15 and that there should be 5 spaces

between "TIME" and "CONC". Proceed as follows: Select sample result output. Then select

and press "E" . Select

"Make new item after this one"

and press "E" two times. Select

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"MOVE TO"

A menu will appear on the screen:

Change item type MOVE TO Change item value 10

Use the numeric part of the keyboard to change the item value to "15". Press "EXIT". You will see a new print selection:

<Left margin of paper or display> 15 <move to this column position> SEQ sequence or tube number

Select

TIME counting time in seconds

and press "E" . Select

"Make new item after this one"

and press "E" two times. Select

MOVE RIGHT move to the right

and press EXIT. A menu will appear on the screen:

Change item type MOVE RIGHT Change item value 10

Use the numeric part of the keyboard to change the item value to 5, then press EXIT.

4.6.9 Printout switches

This selection offers you further choices to modify the printout format. In its initial setting both individual results and average results are printed out. You may switch off or on values shown in the list by selecting NO or YES respectively:

Individual standard values YES Replicate standard averages YES Dilution standard averages YES 2-fields after standards YES Individual unknown values YES Replicate unknown averages YES Dilution unknown averages YES 2-fields after unknowns YES Individual control values YES Replicate control averages YES Dilution control averages YES

Dilution averages effects only if samples are divided into separate dilution groups. 2-fields are those

which would be the same for every item. By means of a switch you can tell the program to only output such fields once after standards or unknowns respectively.

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4.6.10 Copy settings from template

This parameter is to allow you to reuse a particular set o f sample result outputs without having to enter each one individually. You have the choice of selecting the Display sample result output or the output saved in one of three templates:

Single label RIA/IRMA output

i.e. SEQ PAT CODE TIME CPM CONC UNIT %CV FLAG

Dual label RIA/IRMA output

i.e. SEQ PAT CODE TIME CPM CPM_B CONC CONC_B UNIT %CV %CV_B FLAG

Single label RATIO

i.e SEQ PAT CODE TIME CPM

4.7 Leaving the editor

Press "EXIT" to leave the editor A choice of three possibilities is shown:

4.8 Running an assay

Make sure that the first cassette has its correct protocol selection ID and that the appropriate protocol is stored in the instrument. For more information about the " ID system" see section 2.7. Put a "STOP" ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically.

Load racks, starting with the right-hand conveyor lane. Start counting by pressing the START key. The instrument will count all the samples and the RiaCalc WIZ software will evaluate the final results and output

them as determined in the counting protocol. During counting you can see the live results on the live display as described in section 2.4. Alternatively you can

select the menu item "Show evaluation results" to see the output from RiaCalc WIZ on the display. The counting will stop automatically when a "STOP" rack is found. You can also press the STOP key on the

WIZARD keyboard. In that case the following text will appear:

Continue End assay, continue End assay, clear conveyor

Depending on whether you want the next assay to be counted or all counting to stop, select "End assay, continue" or End assay, clear conveyor" respectively. To override the stop instruction and continue counting select "Continue".

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4.9 The FILES function

4.9.1 Introduction

RiaCalc WIZ can produce different types of files. This function in the main menu called FILES allows you to perform operations on and with these files. In CPM mode these files are not produced and in the external MultiCalc mode the same files are handled by MultiCalc on the external computer not on WIZARD.

There are five types of files which can be handled with this function:

Input files Standard curves Results files Controls Trends

In order for anyof these files to be produced you must select the ones you want. You do this with the Saved files parameter in a protocol, see section 4.5.11.

4.9.2 File operations submenu

When you have selected a file a "file operations submenu" will appear. This includes the follo wing items for each file type:

Operation This shows the currently selected operation. If you want a different operation then highlight it and press E. A list of alternative operations will appear. The actual files operation available for each file type are described in later sections.

The actual operation you select will determine what the remaining lines of the file operations submenu are. In every case at least the following lines will appear:

Protocol Selects the protocol to which the file to be operated on belongs. Run id. Specifies the run id. number of the file that is to be operated on. This run id. is selected from a list of free

ids and is not entered with the numeric keys. This is to prevent duplication of run ids. Do operation The submenu for all operations end with this line. Selecting it and pressing E actually starts the

operation defined in the previous lines. If this line is displayed in subdued colour, it means that the selected operation cannot be done.

You can also do the operation by first pressing the EXIT key and then selecting the menu item 'Do specified operation' or alternatively by pressing the Ctrl-S key. In these cases the menu selection bar can be on any menu item.

To return to the main menu without doing the operation first press the EXIT key and then select the menu item 'Quit, do not operate' or alternatively press the Ctrl-C key. You can return to the main menu directly by pressing only the EXIT keyif you have invoked the 'Do operation' and the parameters that specify the operation have not changed after that.

4.9.3 Input file

This comprises data which can be evaluated by RiaCalc WIZ. This data can be either previous output from RiaCalc WIZ or data entered by the user. The main point is that it is data in a format acceptable to RiaCalc WIZ. The functions available for input files are

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Create Creates a new file and lets you edit it. Edit Lets you edit a file. Copy Lets you copy a file. Print Lets you print a file. Evaluate Letsyouevaluateadatafile. Delete Lets you delete a file.

If the selected operation is Create then the line Counting time will appear in the file operation submenu. In principle each measurement result in a data (input) file has its own measurement time. However, in practice the measurement times for all samples in an assay are often the same. When a new data file is created, the counting time given here is assigned to all measurements in that file. This makes the creation of data files easier.

Edit An input data file consists of a series of records, one for each stored measurement result. Each record contains the measurement time, measured counts, CPM's and their errors for one or two channels.

This editor shows only the CPM-values and the measurement time of the first measurement in the file. When the data file is saved, this measurement time is assigned to all records in the file and for each record the counts and error fields are calculated based on the measurement time and the CPM value(s).

You can move up or down one page by pressing the PgUp or PgDn keys respectively, and to the beginning or end of the file by pressing the Home or End keys respectively. To edit a measurement or to jump to a specified line number press the E key.

If you have created a new data file and have not added any CPM's to it yet, try the following shortcut. Use only the E key to move from one menu or menu item to the next appropriate one when you enter new CPM values.

The menu items which may appear during editing are: Make new line after this one You can insert a new line after the currently selected one by i nvoking this menu

item. Chn A cpmEnter here the CPM value for channel A. If CPM fields for both channel A and channel B are empty,

it means that the measured tube was missing.

Chn B cpmEnter here the CPM value for channel B if you want to use it. Cut this line If you invoke this menu item, the line selected for editing is deleted to a "cut buffer". It can be later

pasted by choosing the menu item 'Paste item after this one'. PastelineafterthisoneIf you invoke this menu item, the previously deleted line is pasted fro m the cut buffer

after the line that was selected for editing. Jump to a line If you invoke this menu item, you are asked to give the number of the line to where you want to

jump. Then you must press the EXIT key. When you have finished editing press the EXIT key. In the case of the Copy operation the file operations submenu includes the following additional lines:

Copy to protocol Specifies to which protocol the copy of the file is to be attached. Copy to run id. Specifies the run id number that the copy of the file is to have. If t he protocol is the same as for

the copied file then the run id. must be different. When you access the list of possible run ids. then you will see that used run ids. do not appear on the list so you cannot duplicate them.

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When data has been saved in an input file it can be evaluated again. In addition to editing the data before evaluation you can change the curve fitting method during evaluation if you have selected Halt for curve edit in protocol setting.

4.9.4 Results files

Results files comprise data but they are in ASCII format and are intended for output to an external PC or mainframe which is able to handle the widely accepted ASCII format.

View Lets you view (but not edit) a file. Print Lets you print a file. Send to PC Lets you send a file to PC. Delete Lets you delete a file. Save to disk Lets you save result files in the \EVAL director y of a micro-floppy disk.

When you have a View a result file you can move up or down one page by pressing the PgUp or PgDn keys respectively and to the beginning or end of the file by pressing the Home or End keys respectively.

Note: the view mode only shows the first 40 characters of any line in the Result file and does not allow any editing of the values.

4.9.5 Standard curve files

Standard curves are produced in RIA and IRMA and can be viewed with this function. Note: For standard curve files the run id. designation "REF" means the reference curve.

View Lets you view a file. Delete Lets you delete a file. Set reference curve Lets you set a copy of the selected standard curve as the reference curve for this protocol.

Standard curves can be viewed and if required edited. First you must select the curve you want to view. When it is displayed on screen four functions appear at the bottom of the display:

Edit (1) Press key 1 to start editing. Print (2) Press key 2 to print the curve. Undo (C) Press Clear (backspace) to clear all the changes made in editing and to restore the original curve to the

display: Exit (EXIT) Press EXIT to quit from curve viewing. The editing operations are as follows:

Standard selection. Use the left/right arrow keys to select a particular standard on the curve Replicate selection Use the up/down arrow keys to select a particular replicate point for the currently selected

standard. Delete (1) Pressing 1 deletes the currently selected replicate point. The point which is in the form of a cross will

be replaced by a square. Although this point is still shown on the screen it will not be taken into account in curve fitting. If an inserted point is deleted then it will be taken completely away.

Move (2) Pressing 2 followed by the up/down arrow keys allows you to change the vertical position i.e. response value of the currently selected replicate.

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Insert (3) After pressing 3 put the cursor at the point where you want a new point to be and press E. The new point will be in the form of a cross and will be taken into account in curve fitting.

Undo (C) Pressing Clear (Backspace) will undo the effects of the previous editing. Fit (E) When you press E the existing curve points will be fitted and the selection bar at the bottom of the display

will revert to the Edit, Print, Undo, Exit options. You must use this option to exit from Editing.

4.9.6 Control files

These are produced as part of the quality control operations of RiaCalc WIZ. They can be viewed or deleted with this file function.

Control type This allows you to specify the control type that is to be operated on. In principle there are six types: LOW, MEDIUM, HIGH, CONTROL4, 5 and 6. However the actual types which appear for viewing are only those which were selected in protocol setting.

When you have selected the type and "do operation" the control plot will appear allowing you a number of functions for editing the plot. These functions are the same as for trend plots and are described below.

4.9.7 Trend files

Trend files can also be viewed or deleted after they have been produced. Trend type Specifies the trend type that is to be operated on. This list is fixed because unlike controls it is not

affected by the protocol settings. The options available are:

4.9.8 Control and trend plot editing

The functions available for editing control and trend plots are:

Left/right arrows for selecting the value to be edited. Up/down arrows for defining how many steps to move each time the le ft/right arrow is pressed. WinL(1) Press 1 to set the left limit of a window. WinR(2) Press 2 to set the right limit of a window. Calc(3) Press 3 to recalculate values after making changes to a plot. Del/Undel(4) Press 4 to delete a point. The + mark will change to a square and the point will not be included in

calculations. Pressing 4 when the cursor is on a deleted point will cause the point to be treated again as a normal point.

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Era(5) Pressing 5 allows points within a windowto be permanently removed. They cannot be returned. Print(6) Press 6 to printout the current plot.

When you exit from plot editing (done by pressing E) the changes you have made will be saved.

4.9.9 Spectra

Store spectra - this is for information only. It tells if spectra can be saved or not and if they are to be sent to an

output device or not. The actual settings can be made in SYSTEM | Operation mode | Store assay spectra. See there for more details.

4.9.10 GLP data

This allows you to handle the GLP data obtained in GLP test normalization. There are four options, some of which lead to other options:

View - allows you to view the GLP data Delete - d elete GLP data Criteria - allows you to select the warning limits for different types of G LP data. You can select: Isotope - you can select the isotope t ype from t hose for which a GLP normalization has been done Item - many items appear from which you can select the one which you want the GLP data:

PEAK, BGRD, EFFIC, RESOL, EFFICIENCY, COVERAGE, CHI-PROB, WIN-CPM, TOTAL CPM

Print criteria - select the isotope for which you want the criteria to be printed out.

4.9.11 Waste log file

A waste log file that contains the total CPS and DPS values of all measured assays, isotope normalisations and GLP TEST measurements can be printed or stored on a datalogger disk.

The file can also be deleted or sent via the PC port to an external computer. The waste log can contain approximately 700 entries, after this the older half of the entries is deleted and the log

starts growing again. For isotope normalization and GLP test, TOTAL CPS is the average corrected sample activity in the isotope

For assay measurement TOTAL CPS is the sum of all printed corrected CPM values of measured tubes in the assay converted to CPS. The DPS value is obtained by dividing the printed corrected CPM (converted to CPS) by the actual coverage of the isotope counting window and by the parameter Efficiency% referred to above. TOTAL DPS is thus the sum of all DPS values of measured tubes in the assay.

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4.10 Selectable outputs

MOVE TO Move to this column MOVE LEFT Move to the left MOVE RiGHT Move to the right NEW LINE Move the rest of the text to a new line Vert. Vectors Print sample output result items (e.g. POS, TIME) vertically RACK ### 1 Rack number 3 digits for both groups, no average DET ## 1 Detector number for both groups, no average (always 1 for 1480) SEQ #### 111 Sequence or tube number PAT #### 0,111 Patient number with both averages, unknowns only TIME ##### 1 Counting time in seconds ETIME ####.## 1 Elapsed time in decimal hours COUNT ####### 11 $ Total counts and first average (raw data) COUNT_B ####### 11 $ Total counts and first average on B-channel (raw data) CPM ######.# 11 $ Corrected CPM* and first average CPM_B ######.# 11 $ Corrected CPM* and first average on B-channel CONC ####.### 111 $ Concentration with both averages CONC_B ####.### 111 $ As above for channel B %CV ##.## 011 Coefficient of variation as a percentage, only averages %CV_B ##.## 011 As above for channel B %CVE ##.## 0,01 %CV from reference precision profile, average for only unknowns %CVE_B ##.## 0,01 As above for channel B CODE 111 Code text from coding and control list FLAG " " 0,111 Concentration flag, all values but only unknowns FLAG_B " " 0,111 As above for channel B BLANK ######.# 111 BLANK cpm, valid after BLANK in coding, all values for both groups BLANK_B ######.# 111 As above for channel B REFER ######.# 111 As above for REFER REFER_B ######.# 111 As above for channel B TOTAL ######.# 111 As previously for TOTAL TOTAL_B ######.# 111 As above for channel B NEG ######.# 111 As above for negative controls NEG_B ######.# 111 As above for channel B POS ######.# 111 As above for positive controls POS_B ######.# 111 As above for channel B RESP ######.# 11 $ Programmable response RESP_B ######.# 11 $ As above for channel B STS " " ## 0,111 Numerical flag number. These are:

STS_B " " ## 0,111 Numerical flag number channel B

GROUP "GR" ## 0,11 Group number of multiple UNKN-coding REPL "RP" ## 0,1 Replicate of individual sample for unknowns

SAMPLE "SPL" #### 0,111 Sample No. (includes controls) for all values but only unknowns DRESP #####.# 11 $ Response error DRESP_B #####.# 11 $ Response error for channel B DILF ###.## 0,11 Dilution factor REMARK ############### 0,111 Remark SEQA #### 1 Application sequence ROW ### 111 Row number 1 = individual, 2 = average, 3 = second average DATE ######## 111 Date CLOCK ######## 111 Time CLASS ###.# 111 $ E.g. 1 if CONC = STD 1 or 2.5 if CONC = (STD 3 + STD 2)/2 CLASS_B ###.# 111 $ As above for channel B UNIT (Defined by the user)

1 = OUT, 2 = >STD, 3 = <STD, 4 = ?amb, 5 = >>STD, 6 = <<STD, 7 = %CV!, 8 = >%CV, 9 = %CV>>

* The corrections in CPM are dead time, decay, background, crosstalk and spillover. $ Some fields are used in statistical calculations e.g. a mean or average is calculated. In the above list such fields are marked with a dollar symbol §. # The # marks show the format of the fields. The numbers after fields are the default switch settings. See section 4.6.9 for more details.

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

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5.1 Introduction

The instructions here describe the routine operation of WIZARD when it is connected to a PC running MultiCalc software. Since there are many ways to connect the counter the following list should not be taken as the only possible. In the example a single WIZARD is connected to a PC running MultiCalc laboratory data management software. The operation of MultiCalc is explained in the MultiCalc User Manual.

Note. the MultiCalc communication protocol should be the one designed to work with the version of WIZARD you are using. This will be found on the WIZARD program disk and should be copied from there during installation.

5.2 Start up

1. Switch on the printer. (Note: the printer must be connected to the PC).

2. Make sure that MultiCalc has been installed on your PC. If it has not then follow the instructions in the MultiCalc manual.

To start MultiCalc when the DOS prompt on your PC shows e.g. C: type in the command WIACALC and press Enter. After a short while the PC display will show the MultiCalc main menu.

At the bottom of the PC screen you will see eight softkeys labelled F1 to F8 corresponding to the function keys on your computer keyboard also labelled F1 to F8. A softkey changes its function according to the step in the program you have reached. The actual function of each key at any time is s hown on the bottom of the PC screen. The eight softkeys which have functions in the main menu are:

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F1 COUNTER F2 EVALUATE F3 WORKLISTS F4 PROTOCOLS F5 INP.FILES F6 RESULTS F7 LEVELS F8 ETC.

In addition to the eight softkeys the keys F9 and F10 have fixed functions. Function key F9 is always EXIT and F10 is HELP.

As an alternative to using softkeys there are quick commands listed on the MultiCalc main menu. By pressing the appropriate letter you can make the program go directly to a p articular function instead of by pressing one or more softkeys.

MultiCalc supports different levels of operation e.g. learning or advanced, with automatic or optional helps etc. Choose the level that most suits you. You do this by pressing the softkey F7 (=LEVELS) and then the appropriate LEVEL softkey (F1-F5).

3. Check that microfloppy named 1480 Datadisk is in the WIZARD disk drive. If not, place it there so that the name label is upwards. Switch on WIZARD. After about 3 min. the display will show:

==1480 Main Menu=====================

OPERATE PROTOCOL FILES SYSTEM

==Submenu===========================

Show cpm results

Show evaluationresults

Operate conveyor

==MultiCalc========= ====== === == ==== ======

PressSTARTto measure

Check the time and date by selecting SYSTEM mode and DATE. If it is not correct then give the correct value (See "System | Time & Date setting"). Press EXIT to return to the main display.

Make sure that the counter is in the right mode, i.e. that the text MultiCalc is showing in the lower part of the screen, as in the example here. If it is not, select "SYSTEM" and then the operation mode. The evaluation must be "MultiCalc" .

Now you are ready to start "MultiCalc assay protocol operation" of WIZARD.

5.3 MultiCalc protocol

MultiCalc is controlled by "protocols". A "protocol" is a list of parameters which need to be set or given values e.g. protocol name or counting time.

A MultiCalc protocol is used mainly to define the way the data from the counter is to be handled e.g. items to be output, quality control curves to be plotted etc. These parameters are described in detail in the MultiCalc User

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Manual. The line 3 parameter is called "Measuring parameters". This is where you give the isotope number. See

5.4.3 Isotope selection and 9.30 "Isotopesdefined for 1480 WIZARD". You just select the number of a suitable isotope from the list of those available. Make sure that the one you select has been normalized. If it has not, you need to make one as described in Part 6 Normalization.

5.4 Editing a MultiCalc protocol

Starting from the MultiCalc main menu and depending on whether you want to edit an existing protocol or create a new one you can either press the letters E for protocol edit or F for protocol create or you can press softkey F4 (PROTOCOLS) followed by F1 (EDIT) or F2 (CREATE). You must then select a protocol number from the list of protocols. In the case of EDIT you must select an existing protocol but in the case of CREATE you must give a new protocol and number. Usually t his number will be the next one in the number sequence, but you can give a different number if you want. You must then select the Technology which in this case is Gamma.

If you create a protocol you must then specify the type, RIA, IRMA or RATIO. This choice determines the type of parameter list you get.

The major parameters in the protocol are mentioned below. They are described in detail in the MultiCalc User Manual under "Protocol parameter setting".

5.4.1 Single/Dual label selection

LABEL Select YES if you want to count dual label samples otherwise select NO.

5.4.2 Length of counting

02 COUNTING TIME, MAX COUNTS Give the counting time in seconds and optionally the cut-off value for the number of counts accumulated. A larger count value than this number stops the counting even if the counting time limit has not been reached.

5.4.3 Isotope selection

03 MEASURING PARAMETERS T his is where you select the isotope and the parameters which define measurement using that isotope. When you select parameter li ne 3, three softkeys appear:

F1 I-125 F2 Co-57 F3I+Co

F1 and F2 correspond to isotope numbers 1 and 2. F3 is for dual label counting with iodine and cobalt. Any other isotope must be selected by typing t he number, see the default list in chapter 9 Specifications but also the following section about limitations.

5.4.4 Available isotopes

The used isotope(s) are given in the MultiCalc assay protocol parameter “03 MEASUREMENT PARAMETERS”. We call this parameter here “isotope number”. Its range is 1..99 and it can be interpreted either as one isotope code number in the range 1..99 or as two concatenated isotope code numbers in the ranges 1..9 and

1..10. If you set

DEFINE WIZARD = 1

in communication protocols WIZARD.C00 or WIZARDBG.C00 you can use isotope codes 1..99 in single evaluation assay pr otocols. In dual evaluation assays (having the assay protocol parameter 01 DUAL ASSAY = YES) you can only use i sotopes 1..9; in this case you concatenate the two isotope codes to form a two-digit number. Number ‘3’ is reserved to be equal to ‘12’. (How to measure the isotope number 3 in single label is

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explained in the next paragraph). If in a dual evaluation assay the second digit of the isotope number is 0, it means that the channel B isotope hascode 10.

If you set

DEFINE WIZARD = 0

you can only use isotopes 1..9 in both single and dual evaluation assay protocols. However, even for single evaluation assays you can specify two isotopes by concatenating the two isotope codes to form a two-digit isotope number. In this case WIZARD uses two counting windows and the results can be retrieved during assay evaluation in variables COUNT, CPM and COUNT_B, CPM_B respectively. Single evaluation assays can only use one standard curve, even if two isotopes are used. As be fore, the number ‘3’ is reserved to be equal to ‘12’. To measure the isotope number 3 in single label, use the number 33 and set the assay protocol parameter “01 DUAL ASSAY = NO”.

The communication protocol WIZARD_T.C00 cannot be used to send measurement parameters to WIZARD, so the constant WIZARD does not appear in it. If you are using this communication protocol, set in WIZARD the parameter “SYSTEM | Operation mode | Evaluation” to “CPM” or “RiaCalc WIZ” and edit the assay protocol in WIZARD. You can save the MultiCalc assay protocol on a diskette and then load it into W IZARD. Check, however, that the specified isotope is the right one also after the protocol has been loaded into WIZARD.

5.4.5 Using the communication protocol WIZARD

As described above, this communication protocol is used when resutls are buffered in WIZARD. Each buffered assay is deleted only after MultiCalc has acknowledged that it has received it.

To use this communication protocol, set in WIZARD the parameters “SYSTEM | Operation mode | Evaluation” to “multiCalc” and “SYSTEM | Printout selections |Without buffering to PC” to “No”. If you are not using a local printer that is connected directly to WIZARD, set “SYSTEM | Printout selections |Use printer port” to “No”.

5.4.6 Softkey “F5 INSTALL”

To see what isotopes are available, press in MultiCalc the softkeys “F1 COUNTER | F5 INSTALL”. You get a list of available isotopes and short instructions on how to specify them in assay protocols. (If you have just turned on WIZARD you may get the “Framing error” message at this point. In this case, press ENTER and “F5 INSTALL” again.)

5.4.7 Curve plotting parameters

In the case of a RIA or IRMA the following parameters are available:

20 X-AXIS (concentration) 21 Y-AXIS (response) 22 FITTING ALGORITHM 23 STD OUTLIER REJECTION 24 HALT FOR CURVE EDIT

These are all to do with the standard curve used for determining concentration values. The built-in MultiCalc helps explain these parameter along with the part in the MultiCalc User Manual called Standard curves.

5.4.8 Quality control parameters

The next block of parameters are concerned withquality control. Theyare:

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60 CONTROLS 61 HISTOGRAM 62 QC-ACCEPTANCE RULES

See the MultiCalc Quality Control manual.

5.4.9 Output parameters

The final block of parameters is concerned with output from MultiCalc: 80 STORED FILES These are the types of files stored e.g. data for further analysis with MultiCalc or other

programs or computers, curves, QC information etc. 81 DISPLAY These are the items displayed 82 PRINTER These are the items printed 83 OUTPUT Here you specify what the actual ite ms are in for storing in ASCII format. 84 RESULTS Here you specify what the actual items are in the files for external programs or computers.

5.4.10 Additional output information

The program must know in which order RIA standards are loaded and what their nominal concentrations are. This information is supplied in the CODING accessed by pressing F1. Other special samples such as Blank and Reference can also be specified.

The recommended order for sample tubes is: blanks(NSB)

totals references (zero sample) standard sam ples unknowns.

Unknowns are either patient samples or control samples in arbitrary order, the controls must be further specified on the line called CONTROLS.

2 BLANK 2 TOTAL 2 STD=1 2 STD1=2.5 2 STD1=5 2 STD1=20 2 UNKN

The number on the left is the replicate number and the number on the right is the nominal concentration. The replicate number preceding UNKN means that all unknowns have this number of replicates. In the example above all samples are duplicates and no REFER samples are used.

OtheroutputsavailablewithF2toF7andF8F5andF8F7areCOMMENT,FACTORS,BEGIN,INPUT, OUTPUT OPTIONS, PATIENT and COUNTER.

For more details about MultiCalc assay protocol p arameters see the MultiCalc User Manual "Protocol parameter setting".

A list of available output items for the MultiCalc assay protocol is shown in section 5.8 at the end of this chapter.

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Entry of parameter values in a protocol is supported by various softkeys described in the MultiCalc User manual in the part referred to above.

5.4.11 Exiting assay protocol editing

When you have finished protocol editing you exit by pressing key F9 and then normally selecting F1 (=QUIT+SAVE).

The protocol then joins the list of available MultiCalc protocols. To return to the main menu you must press F9.

5.4.12 MultiCalc protocol operations

If you want to manipulate MultiCalc protocols themselves (rather than just individual parameters in a protocol) e.g. copying a protocol or ordering the list alphabetically etc. you can do this by using the appropriate softkeys (see the MultiCalc User Manual: Introduction to MultiCalc operations).

5.5 Running an assay

5.5.1 Starting the run

Make sure that the firstcassette has its correct protocol selection ID. For more information about this, see "ID system" in Part 2 of this manual. Put a "STOP" ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically.

Load racks, starting with the right-hand conveyor lane. Select the MultiCalc main menu Press "COUNTER", softkey (F1 key) Select 1480 and press"ENTER" MultiCalc will now store all protocols in the instrument automatically and the instrument will count all the loaded

samples.

5.5.2 Stopping the run

The counting will stop automatically when a "STOP" rack or ID is found. You can also press the STOP key on the WIZARD keyboard. In this case the following text will appear on the built-in display:

Continue End assay, continue End assay, clear conveyor

Depending on whether you want the next assay to be counted or all counting to stop select "End assay, continue" or End assay, clear conveyor" respectively. To override the stop instruction and continue counting select "Continue".

5.5.3 Result handling

There are a number of options concerning the way the results are handled. These depend on the type of communication protocol you have. The normal situation is as follows:

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Buffering of results is selected - the SYSTEM parameter "Printout selections | Without buffering to PC" is "NO". If counting is started bypressing the instrument START key or from MultiCalc by pressing F1 COUNTER and selecting WIZARD, measurement results are buffered in t he instrument.

MultiCalc will evaluate the final results and print these out as determined in the counting protocol. Buffering is not selected - "SYSTEM | Printout selections |Without buffering to PC" is "YES". If counting is

started from MultiCalc or by pressing the instrument START key, measurement results are sent directly to MultiCalc, without buffering. This option is recommended only when the Resident filer is installed, in which case you should have the communicationprotocol WIZARD BG instead of the normal WIZARD.

The following special cases are also possible:

1. "SYSTEM | Operation mode | Evaluation" is "RiaCalc WIZ" or "Cpm" and "SYSTEM | Printout selections | Without buffering to PC" is "YES". In this case measurement results are sent directly to MultiCalc, without buffering.

2. "SYSTEM | Operation mode | Evaluation" is "RiaCalc WIZ" or "Cpm" and "SYSTEM | Printout selections | Without buffering to PC" is "NO". In this case measurement results are not sent at all to MultiCalc.

5.6 The FILES function

Most FILES functions are not available in CPM mode, (they are in grey), they are only available in RiaCalc WIZ, however the following three are:

5.6.1 Spectra

Store spectra - this is for information only. It tells if spectra can be saved or not and if they are to be sent to an

output device or not. The actual settings can be made in SYSTEM | Operation mode | Store assay spectra. See there for more details.

are stored using the full 1024 channels. This file is in binary format. If you are only interested in the lower part of spectra, you can set here the number of channels that are included when the file is converted to text form. The channels included always start from the first channel.

Format – When assay spectra are saved in the datalogger disk or are sent to PC or mainframe, you can specify with this parameter the format of the data. Available formats are ‘Wallac’, ‘Excel’ or ‘Ortec’.

- Wallac format: This is a text file that can be read by Wallac Spectrum Analysis Program

- Excel format: This text file format data belonging to the same spectrum is written in the same line and

channel counts values are separated by tabulator characters. This makes it easier to read several spectra at the same time into a spreadsheet program.

- Ortec format: This binary file format is called ‘Integer Data File’ in Ortec documentation. The file extension

is CHN. Each spectrum is stored in a file with the following path and name:

A: \ EVAL \ <assay name>.E<run id number>\<assay position>.CHN

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5 Operation with external MultiCalc

<run id number> can take the values 00, 01, 02, … , 99 and <assayposition> the values 000, 001, 002, …,

999. If “FILES | Spectra | Operation” is “Send to PC”, then all spectra files are sent at the same time one after

another and the r eceiving end must separate them apart.

5.6.2 GLP data

This allows you to handle the GLP data obtained in GLP normalization. There are three options, some of which lead to other options:

Operation:

- View - allows you to view the GLP data

- Delete - delete GLP data

- Criteria - allows you to select the warning limits for different types of GLP data. You can select:

- Print criteria

- Item - many items appear from which you can select the one which you want the GLP data: PEAK, BGRD, EFFIC, RESOL, EFFICIENCY, COVERAGE, CHI-PROB, WIN-CPM, TOTAL CPM

Isotope - you can select the isotope type fro m those for which a GLP normalization can be done Do operation

5.6.3 Waste log file

A waste log file that contains the total CPS and DPS values of all measured assays, isotope normalisations and GLP TEST measurements can be printed or stored on a datalogger disk.

The file can also be deleted or sent via the PC port to an external PC. The waste log can contain approximately 700 entries, after this the older half of the entries is deleted and the log

starts growing again. For isotope normalization and GLP test, TOTAL CPS is the average corrected sample activity in the isotope

For assay measurement TOTAL CPS is the sum of all printed corrected CPM values of measured tubes in the assay. The DPS value is obtained by dividing the printed corrected CPM by the actual coverage of the isotope counting window and by t he parameter Efficiency% refered to above. TOTALDPS is thus the sum of all DPS values of measured tubes in the assay.

5.7 Real time clock

The real time clock of the counter is set from MultiCalc througha command included in the communcation protocol.

If for some reason you want to set the clock manually you can do it by giving one of the following commands when MultiCalc is in terminal mode:

CLOCK dd.mo.yy hh:mi:ss This sets the data and time. To set only the date, send the command

CLOCK dd.mo.yy

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To set only the time, send the comman CLOCK hh:mi:ss

or CLOCK hh:mi

The date is not accepted in any other formats than dd.mo.yy. The year must be a two-digit number. The other values must also be expressed as two-digit numbers, e.g. the month of May is 05.

Values from 80 to 99 refer to the 20th century and values below 80 to the 21st century. The time must be in 24hour format.

After the date has been set, the counter responds with the string DATE SET

After the time has been set, the counter responds with the string TIME SET

If for some reason the date and/or time string could not be interpreted, the counter responds with ? 7 Bad date or time string.

If date or time is set, (as both normally are) the sending of time-of-day values with RIA/IRMA/RATIO assay results to MultiCalc is also enabled.

The exact time when measurement of each sample was started can also be sent to MultiCalc. In MultiCalc mode the measurement start field can be enabled and disabled only from MultiCalc; the SYSTEM

parameter Diagnostic output | Print meas. start time" has no effect in this case. To enable the sending of measurement starting time with assay results, send from MultiCalc terminal the

command CLOCK ON to the counter. To disable it, send the command CLOCK OFF

The sending of this command can be made automatic if it is included in the WIZARD communication protocol by setting the para meter USECLK to 1. See the information that appears when you select F1 COUNTER F5 INSTALL.

The clock value will be assigned to the parameter COUNT_B. This is only available for this use in single label counting.

You can check the current setting by sending the command CLOCK

If the measurement starting time has been enabled, the counter responds with the string PRINTED

If it has been disabled, the counter responds with the message NOT PRINTED

When the counter is turned on or after a power failure in MultiCalc mode, the measurement start time field is disabled until it is explicitly enabled again from MultiCalc.

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5.8 Selectable outputs

RACK ### 1 Rack number 3 digits for both groups, no average DET ## 1 Detector number for both groups, no average (always 1 for 1480) SEQ #### 111 Sequence or tube number PAT #### 0,111 Patient number with both averages, unknowns only TIME ##### 1 Counting time in seconds ETIME ####.## 1 Elapsed time in decimal hours COUNT ####### 11 $ Total counts and first average (raw data) COUNT_B ####### 11 $ Total counts and first average on B-channel (raw data) CPM ######.# 11 $ Corrected CPM* and first average CPM_B ######.# 11 $ Corrected CPM* and first average on B-channel CONC ####.### 111 $ Concentration with both averages CONC_B ####.### 111 $ As above for channel B %CV ##.## 011 Coefficient of variation as a percentage, only averages %CV_B ##.## 011 As above for channel B %CVE ##.## 0,01 %CV from reference precision profile, average for only unknowns %CVE_B ##.## 0,01 As above for channel B CODE 111 Code text from coding and control list FLAG " " 0,111 Concentration flag, all values but only unknowns FLAG_B " " 0,111 As above for channel B BLANK ######.# 111 BLANK cpm, valid after BLANK in coding, all values for both groups BLANK_B ######.# 111 As above for channel B REFER ######.# 111 As above for REFER REFER_B ######.# 111 As above for channel B TOTAL ######.# 111 As previously for TOTAL TOTAL_B ######.# 111 As above for channel B NEG ######.# 111 As above for negative controls NEG_B ######.# 111 As above for channel B POS ######.# 111 As above for positive controls POS_B ######.# 111 As above for channel B RESP ######.# 11 $ Programmable response RESP_B ######.# 11 $ As above for channel B STS " " ## 0,111 Numerical flag number. These are:

STS_B " " ## 0,111 Numerical flag number channel B

GROUP "GR" ## 0,11 Group number of multiple UNKN-coding

REPL "RP" ## 0,1 Replicate of individual sample for unknowns

1 = OUT, 2 = >STD, 3 = <STD, 4 = ?amb, 5 = >>STD, 6 = <<STD, 7 = %CV!, 8 = >%CV, 9 = %CV>>

* The corrections in CPM are dead time, decay, background, crosstalk and spillover. $ Some fields are used in statistical calculations e.g. a mean or average is calculated. In the above list such fields are marked with a dollar symbol, §. # The # marks show the format of the fields. The numbers after fields are the default switch settings.

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6 Normalization

6.1 Normalization

6.2 GLP test normalization

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6.1 Normalization

6 Normalization

6.1 Normalization

6.1.1 Principle

Normalization is a process of optimizing the detector gain and window for each isotope used with WIZARD to ensure the best counting conditions i.e. that the optimum efficienc y and background ratio is achieved.The background must be measured and subtracted in the energy range(s) used. In dual label measurements one isotope being measured will often cause counts to be recorded in the second isotope window and vice versa. This effect is called "spillover" and must be c orrected for.

WIZARD makes all these corrections automatically based on the information obtained during "normalization". This procedure involves measuring a single label sample for each isotope to be used in a particular energy range.

Normalization can be made infrequently. When you have installed WIZARD, normalize it for the isotopes and energy range(s) you are going to be using. Unless the instrument gives a warning asking for a normalization, a time period of six months be fore you redo the normalization is appropriate.

Once you have made a normalization you can then create or edit a normal counting protocol where you can select the isotope you have normalized and use it to count samples.

If the instrument is not normalized for a particular isotope and at some time you try to run an assay with this unnormalized isotope selected, WIZARD will give an error message telling that the isotope is unnormalized and will terminate the assay.

The procedure for doing background and isotope normalization is explained in the following sections.

6.1.2 Background normalization

A background normalization is a method of determining the background for each energy range detector. It is important that it has been made at least once. The background value is subsequently subtracted from count values during actual counting.

6.1.2.1 Background normalization for both energy ranges

To make a background normalization stick the ID label "BKG" onto an ID clip in the area marked "RACK/SPECIAL". Fix the clip onto an empty rack.

Load the rack on the conveyor and press "START". Put a STOP rack after the background rack to stop counting. The background will be measured in both energy ranges and a complete background normalization report will be

printed out and background values for the entire energy spectrum for both energy ranges will be saved in the instrument memory. Since the whole spectrum is saved there is no need to make background measurements for each individual isotope.

6.1.2.2 Background normalization for one energy range

Proceed as in 6.2.1 but include an additional odd or even numeric barcode label in the "PROT OCOL" area of the clip where the odd label selects the normal energy range (15 -1000 keV) and the even selects the extended range (15 - 2000 keV). The background is then only measured in the selected energy range.

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6.1 Normalization

6.1.3 Normalization for isotopes used

Information about the isotopes to be normalized (name, ID number, energy, window settings etc. ) must be given to the instrument before the normalization starts. This information is factory set for the isotopes shown in the list in chapter 8.2 Specifications. Remove the holders from positions 1 to 9 or 1 to 4 (depending on rack type).

Place an isotope source in the last position of an otherwise empty rack. The isotope should have an activity of between 50 000 DPM and 200 000 DPM.

Stick an ID label "NORM" to the ID clip in the position marked "RACK/SPECIAL". Stick an ID label with the appropriate isotope code number to the ID clip in the position marked "PROTOCOL".

The isotope codes are the numbers given in the table in 8.3 Specifications. Fix the clip to the rack and load it onto the conveyor. Insert an empty rack after the last normalization rack to stop

the conveyor. When you have loaded your normalization rack(s) and stop rack, press "START". Repeat this normalization p rocedure for each isotope to be used. Wait until the complete report, for each isotope, has been printed out. Make sure that the efficiency is within the

limits 0.9 ... 1.1.

Make sure you do not leave a background normalization or isotope normalization rack o n the conveyor when you have finished with it otherwise you may start an unwanted new normalization and lose the previous results.

6.1.4 Printout items

The printout column headings are: DET, PEAK CHN*, PEAK DEV%*, RESOL%*, ACTIVITY, COUNTS, WINDOW keV*, LOW, HIGH,

DECAYED ACTIVITY, MEASURED COUNTS, DETECTOR EFFICIENCY, RELATIVE ERROR %, HORROCKS EFFICIENCY*, STANDARD CPM, SIGNIF. LEVEL %

• these onlyappear if extended normalization printout ha s been selected. If a multiple isotope assay (MIA) has standard samples (which are normalized at the beginning of the assay) and

they have "SYSTEM  Isotopes  <isotope name>  Norm. zero time" equal to "Start", then the decay correction is done to the beginning of the whole multiple isdotope assay. This makes it possible to compare the STANDARD CPM value calculated during isotope normalization with MIA unknown CCPM values.

6.2 GLP test Normalization

6.2 GLP test normalization

6.2.1 Introduction

Instrument performance can be monitored by running GLP test normalizations at regular intervals. These store data that can later be viewed in graphical format.

GLP means "Good Laboratory Practice". A GLP test normalization is similar to isotope normalization, only results are stored differently. Data obtained in a GLP test normalization are not used in assay measurements, but are tested against preset limits and then stored so that they can later be compared with other test normalizations using the same isotope. This comparison is done by presenting the values of some measured parameters as a function of time, so that any systematic trends or large random deviations can easily be discerned.

6.2.2 GLP test normalization rack

A GLP test normalization rack has only one holder and sample which is at the last position of the rack. The rack has a clip with the TEST instruction at the RACK/SPECIAL position and the isotope code at the PROTOCOL position. Counting time is set by the parameter Normalization time which is found in the SYSTEM menu under "Isotopes | <Isotope name>". The printout is similar to isotope normalization printout.

It is possible to do GLP test measurements by only using the isotope numbers 4, 91, 92 and 93. (If needed, it is also possible to make other isotopes available for GLP test measurements).

6.2.3 Saved GLP values

The following values are saved during GLP test normalization: PEAK - Isotope main peak channel number BGRD - Background CPM in counting window RESOL - Detector resolution (%) EFFICIENCY - Absolute detector efficiency. This is determined for I-125 using the Horrock's method. For other

isotopes, the measured CPM in the counting window is divided by the absolute activityof the test sample, which is given by the SYSTEM parameter "Isotope | <Isotope name> GLP test sample DP M".

COVERAGE - Window coverage (%). This is the fraction of counts in the whole spectrumthat fallsto the isotope counting window.

CHI-PROB - Detector stability probability. This can be calculated if the SYSTEM parameter "Isotope | <Isotope name> Repeat times" is greater than 1. See section 6.2.7 "Repeat Counting" for more details.

WIN-CPM - Measured CPM in counting window TOTAL CPM - Measured total CPM in the whole spectrum

6.2.4 GLP Criteria

For each of the above quantities you can set a low and a high limit by setting the FILES menu item "GLP data | Operation" to "Criteria" and pressing the ENTER key when the highlight bar is on the menu item "Do operation". The other items in the menu are used to select the isotope and one of the stored values mentioned above. In the same menu that is used to set the limit values you can specify that a warning message, a graph or both are printed if during GLP test measurement some quantity is not within limits.

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6.2 GLP test normalization

6.2.5 Viewing GLP data

The stored GLP test normalization data can later be viewed graphically by setting the FILES menu item "GLP data | Operation" to "View" and pressing the Enter key when the highlight bar is on the menu item "Do operation". The other items in the menu are used to select the and one of the stored values mentioned above.

6.2.6 Outputting GLP data

GLP plots can later be printed by pressing the digit key "6" while the plot is displayed. The plot is sent to the printer that is connected to the WIZARD printer port if the SYSTEM parameter "Printout selections | Use printer port" is "Yes".

The plot is also sent via the WIZARD PC port to MultiCalc if either "SYSTEM | Operation mode | Evaluation" is "MultiCalc" or "SYSTEM | Printout selections | Without buffering to PC" is "Yes".

When the plot is sent to MultiCalc, special code characters are added to the data, so that the plot can be printed by the printer that is connected to the PC running MultiCalc. In order for the printing to succeed, MultiCalc must be receiving data from WIZARD, and in the WIZARD communication p rotocol the Terminal parameter must be VT-

52.

6.2.7 Repeat counting

During GLP test normalization each measurement can be repeated several times to test the detector for stability. The parameter "Repeat times" in isotope editor in the SYSTEM menu sets the number of times each isotope

normalization measurement is repeated. Thus the total time a sample is measured in the detector is this number multiplied with the normalization time that is set in the isotope editor.

The measured counts in repeat measurements are compared with each other and the program calculates the probability that differences between expected and observed counts in these measurements occured just because of statistical variation. This probability is called "Significance level" and its unit is %. If it is near zero or one, this means that there i s systematic error in repeat measurements.

The number stored is transformed from the Chi-square probability that is shown in the printout ("SIGNIF. LEVEL") so that

5 corresponds to 50%, 4 to 10%, 3 to 1%, 2 to 0.1%, 1 to 0.01%, 0 to <=0.001%, 6 to 90%, 7 to 99%, 8 to 99.9%, 9 to 99.99%, and 10 to >=99.999%. This is to make very small and large probability values stand out more clearly.

Possible isotope decay is taken into account when Si gnificance level is calculated. The counting window over which counts are summed is the same for all repeat measurements and is determined from the sum spectrum of the repeat measurements.

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7 Additional WIZARD

7.1 Dual label counting

7.2 Multiple-isotope assay counting

7.3 System mode

7.4 High activity mode

7.5 Power failure

7.6 Routine maintenance



functions

7.7 Safety information

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7.1 Dual label counting

7 Additional WIZARD functions

7.1 Dual label counting

7.1.1 Introduction

Dual labelled samples have actually two independent analytes in the same vial. In order to separate the radioactive labels from each other the labels must have separate energies. The most common dual label assay in practical work is the B12/Folate assay in which labels are Co-57 and I-125.

Before running a dual label assay you must normalize the instrument for both isotopes as described in Part 6. The analysis for dual labelled assays requires two protocols to be set, one for each isotope used. The two protocols

are connected by specifying in protocol A the name of protocol B. Protocol "A" is a master protocol and "B" is the slave.

Channel A protocol

Refers to channel B protocol

Figure showing relationship between protocols in dual label. The program uses the word "channel" to describe the counting process controlled by a particular protocol.

Channel A counting is controlled by protocol A and channel B by protocol B. Each channel may include counts from both labels so t he dual label program has to "disentangle" the counts so as to arrive at a pure counts value for each label.

Protocol B must be defined before A otherwise you cannot select the B protocol when you edit the A protocol. The explanation following applies to both the internal RiaCalc WIZ software and the external MultiCalc software.

Channel B protocol

7.1.2 Setting dual label protocols

The following example shows setting of the protocol for a dual labelled B12/Folate assay Create a protocol with the name B12 and ID 11. Answer YES to the question DUAL LABEL. Two extra lines appear, e.g.:

Channel = A Chn-B Protocol = B12

Select:

Channel = B

The chn-B line will disappear. Give the coding for B12 standards and controls as in a single label assay protocol. Select the label:

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7.1 Dual label counting

First Isotope I-125 Second Isotope Co-57

Give the rest of the parameters, including time and coding for the Folate standards and controls as in the case of a single labelled protocol.

Exit and Save the protocol. Make a copy of this protocol with the name Folate and ID 10 Answer YES to question DUAL LABEL Two more lines appear: Select:

Channel = A Chn-B Protocol = B12

Make other necessary changes to e.g. coding. Note: The protocol types for A and B can be different, i.e. one can be RIA and the other IRMA. Exit and Save the protocol. Run the assay as for single label. The protocol ID, which in the example is 10, can be from 1 .. 99.

7.1.3 Parallel and successive evaluation

The actual evaluation may take place in two different forms, parallel and successive. Parallel processing is when results from channel A and B are calculated simultaneously in real time. Successive processing is when results from channel A are calculated and saved in real time. These results are then

retrieved and the results for channel B are calculated later. Parallel processing imposes strong constraints on the assay. Both analytes must have the same number of

standards and controls and the number of replicates must be the same. Successive processing has practically no constraints at all. The number of standards, controls, replicate etc. is

freely selectable. The program selects, according to the protocol, whether the evaluation is parallel or successive.

7.1.4 Constraints on protocol setting

The following is a s ummary of protocol commands and how these are constrained by the requirements for dual label protocols.

DUAL LABEL, COUNTING TIME, LABELS Only the channel A settings are valid. The channel B settings have no effect

X-AXIS, Y-AXIS, FITTING ALGORITHM, STD OUTLIER REJECT, CURVE EDIT HALT May be set differently for channel A and channel B protocols

CONTROLS Must be set identically for both protocols if parallel processing is to occur

PRINTER Can be set differently if successive processing is to be done, but if parallel processing is done then t he channel A settings dominate.

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7.1 Dual label counting

CODING For parallel p rocessing CO DING must be identical, except for the numerical values for concentrations of standards. Unknowns must begin from the same position number and must have similar structure, number of replicates and dilutions (with the same dilution factors).

Hint: If you want to be sure that the result calculation happens in parallel, create the channel B protocol first, edit it and save it. Next, make a copy of it and give it the name channel A protocol. Finally make the necessary changes to the channel A protocol (e.g. add to the channel A protocol the reference in the channel B protocol). Note that the channel A protocol parameter LABELS must include both labels.

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7.1 Dual label counting

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7.2 Multiple-isotope assay counting

7.2 Multiple isotope assay counting

7.2.1 Introduction

The WIZARD multi-isotope assay counting mode (MIA) allows measurement of samples labelled with multiple isotopes in the range 10 - 2000 keV. Results can be corrected both for spillover from one counting window into another and for radioactive decay of the isotopes used for labelling the samples. Two formats are possible for the printout of results to enable them to be displayed in the most convenient way.

The MIA mode is part of the RiaCalc WIZ software package. The number of isotopes you can use depends on the separation of the isotope peaks and hence on the isotopes

selected. The software can handle spillover correction for up to 20 isotopes. At typical multi-isotope spectrum with good separation of the peaksis shown in the figure below.

Normalization of the isotopes used takes place as part of the MIA itself. The standards are loaded into the first rack(s) before the racks with unknown samples. In protocol parameter setting you specify the isotopes used and the order in which the standards are arranged.

If a normalization has already been done you do not need to repeat it. In the protocol, specify the isotopes in the correct order but set the replicate number to zero for those isotopes for which the normalization has already been done.

When you are loading samples, put the standards for the unnormalized isotopes at the beginning of the first rack. You do not need to leave empty positions for those isotopes which are already normalized. WIZARD looks after the arrangement automatically, based on the information in the protocol.

7.2.2 Protocol editing

Create a new MIA protocol or edit an existing one. To create a new one, give the name and protocol number. For the assay type select MIA. Parameter editing is the same as with other assay types but the lis t of parameters available is different. The parameters are as follows:

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--Edit protocol---------------------------------------17 PROT18 MIA

___________________ ___ ___ ___ ___ ___ ___ __ Unkn counting time. 60 Max counts limit 9999999 LCR time 7 LCR counts 10 Unkn replicates 2 Subtract background YES Spill correction YES Printout Standards

--Integer, use number keys---------------------

7.2.2.1 Unkn. counting time

This is the counting time used to measure unknown samples. The unit is the second.

7.2.2.2 Max. counts limit

This parameter specifies the counts value that, when exceeded in all windows of the sample being currently measured, determines that t he measurement be ended. For example, if the required statistical accuracy for all samples is 0.1%, you can set the max. counts limit to 1000000. (You must make sure thatthe countingtime is long enough for the required counts limit to be reached.) The upper value for the max. counts limit is 99 999 999

7.2.2.3 Low Count Reject time

This is the time at the beginning of unknown sample measurement during which it is determined whether the sample is active enough that it is worth while to continue measurement. It is continued if at least the number of counts given by the parameter "LCR counts" has been collected in some counting window during the "LCR time". The unit of "LCR time" is the second.

The Low Count Reject feature is disabled if "LCR time" is greater than the measurement time or if "LCR counts" is zero.

7.2.2.4 Low Count Reject counts

After the "LCR time" has passed since the measurement of a tube was started, a check is made to ensure that at least one counting window has the minimum number of counts given in "LCR counts". If this is not the case, the measurement of this tube is terminated.

7.2.2.5 Unkn. replicates

This is the replicate number for unknown samples.

7.2.2.6 Subtract background

This is used to select whether background is subtracted or not.

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7.2 Multiple-isotope assay counting

7.2.2.7 Spill correction

This is used to select whether corrected counts and CPM are corrected for spillover. In most cases spillover correction should be selected.

7.2.2.8 Printout

This is used to select what result data is printed. The possibilities are as follows (see the figure below):

--Edit protocol---------------------------------------17 PROT18 MIA _Printout______________ ___ ___ ___ ___ ___ __

Horiz. layout No Print replicates Yes Print replicate averages Yes Sequence number Yes

Sample number Yes

Replicate number Yes Elapsed timein hours No Counting timein seconds No Dead time No Counts Yes

--Choice, use

←→ keys --------------More ↓ -----

7.2.2.8.1 Horiz. layout

There are two layouts for the printout. If you select the parameter Horizontal layout to be YES, the printout will appear as below (see also the first printout e xa mple at the end of this chapter):

POS SAMPLE CODE ETIME CTIME DTIME%

Is 1 Is 2Is 3Is 4 COUNTS CPM BGRD ERR% CCPM

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7.2 Multiple-isotope assay counting

CCPM% FCCPM RATIO FACTOR ….. If you select Horizontal format to be NO, the printout has the following format: POS SAMPLE CODE ETIME CTIME DTIME%

COUNTS CPM BGRD ERR% CCPM ....

Is 1 Is 2 Is 3 Is 4

......

7.2.2.8.2 Print replicates

Print results for each individual tube in a replicate set.

7.2.2.8.3 Print replicate averages

Print the replicate average results of a replicate set of tubes. Note: at least one of t he two parameters above must be YES otherwise no results will be printed.

7.2.2.8.4 Sequence number

The numbering for sequence number starts from the beginning of the assay and includes the standard tubes. E.g. if you have five standards the first sequence number printed will be 6. See the examples at the end of this chapter.

7.2.2.8.5 Sample number

The number of the replicate group (or patient) to which the measured tube belongs.

7.2.2.8.6 Replicate number

This is position of a sample in a replicate group. E.g. if you have replicates 3 this "replicate number" will be 1, 2 or 3 depending on the measured tube.

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7.2.2.8.7 Elapsed time in hours

Time since the start of the current assay.

7.2.2.8.8 Counting time in s econds

The time for which t he tube was measured.

7.2.2.8.9 Dead time

Dead time as a percentage of count time.

7.2.2.8.10 Counts

Measured counts in a counting window.

7.2 Multiple-isotope assay counting

7.2.2.8.11 Background

Background counts in a counting window. This is output only if background subtraction is selected.

7.2.2.8.12 CPM

The counts per minute value measured in a counting window. It includes the following corrections: Dead time Background (if selected)

7.2.2.8.13 Error (%) in CPM

Relative error in the CPM value due to statistical counting error.

7.2.2.8.14 Corrected CPM

The corrected counts per minute value. The following corrections are made if specified in the MIA and SYSTEM/isotope protocols respectively:

Spill correction Isotope decay correction

7.2.2.8.15 DPM

The DPM (disintegrations per minute) is obtained by dividing the corrected CPM by a factor comprising the percentage of the counts in the window used and the detector efficiency (these two together make up the counting efficiency).

7.2.2.8.16 RATIO

The relative amount of isotope present in an unknown sample as compared to the standard sample for this isotope.

7.2.2.8.17 CCPM / DPM

This is the "counting efficiency" parameter referred to in DPM above. It is the ratio of the observed counts to the total disintegrations in the same time.

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7.2 Multiple-isotope assay counting

7.2.2.9 Standards

This list is used to specify standard samples at the beginning of a MIA assay. The number on the le ft is the replicate number, then follows the isotope code and name. If you set the replicate number to 0 then a stored normalization will be used and no standard for that isotope should be put in the standards rack.

Note: it is recommended that normally standards will used rather than stored normalizations. To edit, create or delete a line in the list press the ENTER key. You can also delete a line by pressing the DEL key

on a separate PC-compatible keyboard and change the replicate number with the + and - keys.

7.2.3 Leaving the editor

Press "EXIT" to leave the editor A choice of three possibilities is shown:

7.2.4 Running a multi-isotope assay

Make sure that the first cassette has its correct protocol selection ID and that the appropriate MIA protocol is stored in the instrument. For more information about the "ID system" see section 2.1.5. Put a "STOP" ID on the last sample rack to be counted or use a STOP rack or a totally empty rack in order to stop the instrument automatically.

Load racks, starting with the right-hand conveyor lane. Begin with the rack containing the standards (if any). Start counting by pressing the START key.

The instrument will count all the samples and the RiaCalc WIZ MIA software will evaluate the final results and output them as determined in the countingprotocol.

The counting will stop automatically when a "STOP" rack is found. You can also press the STOP key on the WIZARD keyboard. In that case the following text will appear:

Continue

End assay, continue

End assay, clear conveyor

7.2.5 Error message

The error message ”Results for CCPM, DPM and RATIO may be inaccurate" might appear. It means that windows have been set or isotopes selected in such a way that two windows are almost totally overlapping and it is not possible to resolve t he peaks. Change your window settings or isotope selection.

7.2.6 Outputting results to disk

If you want to transfer your results to a disk, e.g. to use with another program, follow the instructions in System mode, section 7.3.6.5.

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7.2 Multiple-isotope assay counting

7.2.7 Printout examples

Examples of the two printout formats are shown on the next four pages. Normalization was done with five standards but the samples were only dual labelled so the numerical values show significant counts only for I-129 and Co-57. The negative CCPMs for the unrepresented isotopes are due to the normal statistical variation in the counts. The program shows the size of this variation by printing the negative results rather than truncating them to zero.

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7.2 Multiple-isotope assay counting

Example of a multi-isotope printout when horizontal layout YES is selected

MULTI-ISOTOPE ASSAY 4 MIA Seven labels -- UNKNOWN SAMPLES 16-Jan-1994 11:23:14

Time limit 60 Max. count limit 9999999 Low count time 6 Low count limit 10 Replicates 1 Subtract background YES Eliminate spillover YES

Sequence number 1 Sample number 1 Replicate number 1 Elapsed time 0.01 hours Counting time 60 seconds Dead time 0.173 %

ISOTOPE Cd-109 Ce-141 In-114m Cr-51 Sn-113 Sr-85 Sc-46 COUNTS 13941 24 23 25 17 19 19 BGRD CPM 7.0 15.0 11.2 24.0 14.0 21.0 19.0 CCPM 13957.4 -4.3 3.1 0.8 -1.6 -7.1 -0.6

Sequence number 2 Sample number 2 Replicate number 1 Elapsed time 0.12 hours <F255D> Counting time 60 seconds Dead time 1.000 %

ISOTOPE Cd-109 Ce-141 In-114m Cr-51 Sn-113 Sr-85 Sc-46 COUNTS 804 67801 867 82 14 29 17 BGRD CPM 7.0 15.0 11.2 24.0 14.0 21.0 19.0 CCPM 23.2 68456.9 7.9 -11.8 -9.3 10.0 -5.6

Sequence number 3 Sample number 3 Replicate number 1 Elapsed time 0.14 hours Counting time 60 seconds Dead time 1.194 %

ISOTOPE Cd-109 Ce-141 In-114m Cr-51 Sn-113 Sr-85 Sc-46 COUNTS 31155 3242 22225 4113 3618 6058 2286 BGRD CPM 7.0 15.0 11.2 24.0 14.0 21.0 19.0 CCPM 785.0 58.3 22375.4 46.0 102.6 303.0 260.9

Sequence number 4 Sample number 4 Replicate number 1 Elapsed time 0.16 hours Counting time 60 seconds Dead time 0.134 %

ISOTOPE Cd-109 Ce-141 In-114m Cr-51 Sn-113 Sr-85 Sc-46 COUNTS 193 463 323 9836 231 22 17 BGRD CPM 7.0 15.0 11.2 24.0 14.0 21.0 19.0 CCPM 11.8 -46.5 -80.5 9831.6 27.5 10.2 8.5

Sequence number 5 Sample number 5 Replicate number 1 Elapsed time 0.18 hours Counting time 60 seconds Dead time 1.015 %

PerkinElmer Wallac 1480 WIZARD 3 User manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Warning

Contents

Trademarks

1 Introduction

1.1 Introduction to PerkinElmer Life and Analytical Sciences

1.2 Introduction to 1480 WIZARD

1.3 Introduction to this manual

2 WIZARD controls

2.1 Introduction

2.2 Keyboard

2.3 Display

2.4 Live display

2.5 Racks

2.6 Principle of the ID system

2.7 Fitting ID labels

2.8 Loading racks the right way round

2.9 Instruction labels

2.10 Numerical labels

2.11 Barcode errors

2.12 Help

3 CPM operation

3.1 Introduction

3.2 Start up

3.3 Normalization

3.4 Protocol editing

3.5 Parameters available

3.6 Output

3.7 Leaving the editor

3.8 Running an assay

3.9 The FILES function

4 Operation with internal RiaCalc WIZ

4.1 Introduction

4.2 Start up

4.3 Normalization

4.4 Protocol editing

4.5 Parameters available

4.6 Output editing

4.7 Leaving the editor

4.8 Running an assay

4.9 The FILES function

4.10 Selectable outputs

5 Operation with external MultiCalc

5.1 Introduction

5.2 Start up

5.3 MultiCalc protocol

5.4 Editing a MultiCalc protocol

5.5 Running an assay

5.6 The FILES function

5.7 Real time clock

5.8 Selectable outputs

6.1 Normalization

6 Normalization

6.2 GLP test Normalization

7.1 Dual label counting

7 Additional WIZARD functions

7.2 Multiple isotope assay counting