Hoefer SP-2001 User Manual

Vision Spectrophotometer
USER MANUAL
Declaration of Conformity
This is to certify that the Vision UV/Visible Spectrophotometers part numbers:
SP-2001, SP-2001PT, SP-2001BL and SP-2001SD manufactured by Hoefer, Inc. conform to the requirements of the following Directives:
73/23/EEC & 89/336/EEC& IVD
Standards to which conformity is declared
EN 61010-1: 2001 Safety requirements for electrical equipment for measurement, control and laboratory use.
EN 61326-2.3: 1998 Electromagnetic compatibility - generic emission standard Electrical equipment for measurement, control and laboratory use.
EN 61000-4-6: 1992 Electromagnetic compatibility - generic immunity standard part 1. Residential, commercial and light industry.
BS EN 591:2001 Instruction for use for in vitro diagnostic instruments for professional use.
BS EN 13612:2002 Performance evaluation of in vitro diagnostic medical devices
2002/96/EC This appliance is marked according to the European directive 2002/96/EC on Waste Electrical
and Electronic Equipment (WEEE). By ensuring this product is disposed of correctly, you will help prevent potential negative consequences for the environment and human health, which could otherwise be caused by inappropriate waste handling of this product.
The symbol
on the product, or on the documents accompanying the product, indicates that this appliance may not be treated as household waste. Instead it shall be handed over to the applicable collection point for the recycling of electrical and electronic equipment. Disposal must be carried out in accordance with local environmental regulations for waste disposal.
Signed:
John Attwood Director of Sales & Marketing Hoefer, Inc.
Hoefer, Inc. 84 October Hill Road Holliston, MA 01746
Toll Free: (800) 227-4750 Phone: +1 508-893-8999 Fax: +1 508-429-5732 E mail: support@hoeferinc.com Website: www.hoeferinc.com
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TABLE OF CONTENTS
ESSENTIAL SAFETY NOTES .................................................................................................................................... 4
Unpacking, Positioning and Installation .................................................................................................... 4
INTRODUCTION.......................................................................................................................................................... 5
Your spectrophotometer .............................................................................................................................. 5
Sample handling tips.................................................................................................................................... 5
Keypad and display ...................................................................................................................................... 6
Software style................................................................................................................................................ 8
THE APPLICATIONS FOLDER .................................................................................................................................. 9
1: Single Wavelength – Abs and %T ......................................................................................................... 10
2: Concentration.......................................................................................................................................... 12
3: Wavescan ................................................................................................................................................ 14
4: Simple Kinetics ....................................................................................................................................... 17
5: Standard Curve ....................................................................................................................................... 20
6: Multiple Wavelength ............................................................................................................................... 24
7: Absorbance Ratio ................................................................................................................................... 25
THE LIFE SCIENCE FOLDER .................................................................................................................................. 27
1: DNA................................................................................................................................................... 29
2: RNA................................................................................................................................................... 31
3: Oligo .................................................................................................................................................. 33
Protein Determination ................................................................................................................................ 35
1: Protein UV ......................................................................................................................................... 36
2: BCA ................................................................................................................................................... 38
3: Bradford............................................................................................................................................. 41
4: Lowry................................................................................................................................................. 44
5: Biuret ................................................................................................................................................. 47
Bacterial Cell Culture Measurement (OD600) .......................................................................................... 50
FAVORITES AND METHODS FOLDERS ................................................................................................................ 52
UTILITIES FOLDER .................................................................................................................................................. 53
Utilities ......................................................................................................................................................... 54
1: Date and Time................................................................................................................................... 54
2: Regional ............................................................................................................................................ 54
3: Printer................................................................................................................................................ 54
4: Preferences ....................................................................................................................................... 55
5: Contrast............................................................................................................................................. 55
6: Folder Names.................................................................................................................................... 55
7: About ................................................................................................................................................. 56
8: Games............................................................................................................................................... 56
ACCESSORIES INSTALLATION ............................................................................................................................. 58
Printer installation ...................................................................................................................................... 58
Loading / changing the printer paper ....................................................................................................... 60
Bluetooth accessory installation............................................................................................................... 61
PRINT VIA COMPUTER............................................................................................................................................ 64
ACCESSORIES ......................................................................................................................................................... 65
Lamp Replacement..................................................................................................................................... 65
Cleaning and general care of the instrument........................................................................................... 65
SPECIFICATION AND WARRANTY ........................................................................................................................ 66
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ESSENTIAL SAFETY NOTES
There are a number of warning labels and symbols on your instrument. These are there to inform you where potential danger exists or particular caution is required. Before commencing installation, please take time to familiarise yourself with these symbols and their meaning.
Caution (refer to accompanying documents). Background color yellow, symbol and outline black.
Unpackin g, Positioning and Installation
Check the contents of the pack against the packing list. If any shortages are discovered, inform your supplier immediately.
Inspect the instrument for any signs of damage caused in transit. If any damage is discovered, inform your supplier immediately.
Ensure your proposed installation site conforms to the environmental conditions for safe operation:
Indoor use only. Temperature range 5°C to 35°C. Note that if you use the instrument in a room subjected to extremes of temperature change during the day, it may be necessary to recalibrate (by switching off and then on again) once thermal equilibrium has been established (2-3 hours). Maximum relative humidity of 80% up to 31°C decreasing linearly to 50% at 40°C
The instrument must be placed on a stable, level bench or table that can take its weight (< 4.5 kg) so that air can circulate freely around the instrument.
This equipment must be connected to the power supply with the power cord supplied. It can be used on 90 – 240 V, 50-60 Hz supplies.
If the instrument has just been unpacked or has been stored in a cold environment, it should be allowed to come to thermal equilibrium for 2-3 hours in the laboratory before switching. This will prevent calibration failure as a result of internal condensation.
Switch on the instrument via the keypad ( series of self-diagnostic checks.
Please read through this user manual prior to use.
Please contact your original supplier in the first instance if you experience technical or sample handling
difficulties.
If this equipment is used in a manner not specified or in environmental conditions not appropriate for safe operation, the protection provided by the equipment may be impaired and instrument warranty withdrawn.
) after it has been plugged in. The instrument will perform a
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INTRODUCTION
Your spectrophotometer
Your spectrophotometer is a simple-to-use UV/Visible instrument with a CCD array detector (1024 pixels). It has no moving parts, which is the basis of the rapid scanning operating system.
The user interface is built around folders which are displayed on the home page when the instrument is switched on. After switch on and calibration, the default home page is "Vision" offering the choice of
Applications General spectroscopic methods
Favorites A folder to store your more frequently used configured methods
Methods Contains nine folders that can store less frequently used configured methods (nine
methods per folder)
Utilities Instrument set up (date, time, language, etc) and games
Life Science Standard Life Science methods such as nucleic acid assays, protein assays and cell
counting
The instrument is supplied with a program PVC (Print via Computer) on the accompanying CD. When used with a USB cable to connect to a PC onto which the software has been installed, it enables the user to “print through” the PC directly to the printer that is connected to it. The data may also be stored as an Excel spreadsheet, as an EMF graphics file, a comma delimited (csv) data file, a tab delimited (txt) data file or in native PVC format for later access
Alternatively, results may be sent to the PC via a Bluetooth accessory; this can either be supplied pre-installed or is available as an optional accessory if the need for its use arises after installation of the product. PVC works in a similar way.
A printer is available for the instrument; this may either be supplied pre-installed or is available as an optional accessory if the need for its use arises after installation of the product.
Sample handling tips
Note that the light beam is directed from RIGHT to LEFT through the cell chamber; therefore please ensure the cell is inserted in the correct alignment.
The cell holder supplied with the instrument accepts standard 10 mm pathlength quartz, glass or plastic cells.
The optical beam height is 15 mm, and the minimum volume that can be used is approximately 10μl in a
Quartz ultra-micro cell.
12 mm test tubes may be used (e.g. for cell cultures), however they are not recommended as higher quality data is produced by using disposable cuvettes for the analysis. If used, align the indicator line on 12 mm test tubes in the same direction to ensure reproducible positioning of the tube. Note that test tubes do not last forever, and that the surface becomes scratched and blemished through repetitive use; if this is the case they should be replaced.
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A
Keypad and display
The back-lit liquid crystal display is very easy to navigate around using the alphanumeric entry and navigation arrow keys on the hard wearing, spill proof membrane keypad.
Key
On/off key Turns the instrument on/off
Arrow keys Use the four arrow keys to navigate around the display and select the required
View Options: ::; View options for that application mode. Some of these are common to all
Alphanumeric keys Use these to enter parameters and to write text descriptions where
Escape/Cancel:
Set Reference: 0A/100%T Set reference to 0.000 A or 100%T on a reference solution at the current
Enter:
Enter, or confirm, a selection. Take a measurement.
Escape from a selection and return to the previous folder. Stop making
ction
setting from the active (highlighted) option.
applications and described below. Options unique to an application are described in the relevant section.
appropriate, or required. Use repeated key presses to cycle through lower case, number and upper case. Leave for 1 second before entering next character. Use C button to backspace and 1 to enter a space.
measurements.
wavelength in the mode selected. When in scan mode, do a reference scan.
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Options (select using key pad numbers)
1. View parameters for the experiments.
2. Print the results. 3,4,5,6 Described in the application.
7. Define the sample number you wish to start from.
8. Save the parameters as a method to a defined folder name with a defined method name.
9. Toggle auto-print on/off. Default is off.
Exit options by pressing
Experienced operators can use the numeric keys as a shortcut to the option required without needing to enter the Options menu.
, or wait.
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Software styl e
The user interface is built around having folders of files which are displayed on the home page when the instrument is switched on. Different folders are numbered and opened by using the associated number key on the keypad.
Summary Function Keypad number Description
1 Single wavelength, Concentration, Wavelength scan, Kinetics, Standard
Curve, Multiple wavelengths and Ratio
2 Saved User selected and configured methods
3 Sub folder selection for user selected and configured methods
4 Instrument set up (date, time, language, etc) and Games
5 Nucleic acids, Proteins and Cell counting
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THE APPLICATIONS FOLDER
SUMMARY:
Function Key pad number Description
1 Absorbance or %T (Transmission) at a single user defined
wavelength.
2 Concentration measurement at a single wavelength based on a
simple Factor entered or calculated from a single standard.
3 Wavelength scan between two user defined wavelengths. Range
200-950 nm, with user configurable peak finding function.
4 Absorbance versus time measurements either rate or end value
based.
5 Generation of calibration curve by measuring standards at a single
wavelength.
6 Absorbance or %T (Transmission) at up to 5 user defined
wavelengths.
7 Ratio of absorbance values at two user specified wavelengths.
OPTIONS
Within each application the user has the possibility to select various options that define the way results are treated. If not using a stored method, it is advisable to check that these Options have been appropriately set for your experiment when coming to the instrument. Note that setting the “History” parameter to on (see Preferences later) will cause the instrument to store it’s last settings. If the “History” parameter is turned off, all parameters and options will return to their default settings when you leave that application. (Unless it has been saved as a method).
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1: Single Wavelength – Abs and %T
This makes simple Absorbance (A) and % Transmission (%T) measurements on samples, measuring the amount of light that has passed through a sample relative to a reference (this can be air). The procedure is as follows:
Step 1
Set wavelength by using keypad numbers or left and right arrows. Press the down arrow key.
Step 2
Select the mode, Absorbance or %T, using the left and right
arrows.
Step 3
To enter the results screen with the selected parameters press
OK OR Cancel the selections and return to the Applications Folder by
pressing Cancel
Step 4
Insert the reference. Press 0A/100% key. This will be used for all subsequent samples until changed.
Step 5
Insert sample and press
Repeat step 5 for all samples.
Results
The result at the selected wavelength is displayed on screen. Use the left and right arrows to move the cursor and display the value at the cursor position (+/- 15nm from set wavelength).
Press Cancel
Press ::; to display available Options which are described below.
.
.
to return to the Applications Folder.
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Options (select using key pad numbers)
1. Return to parameters screen (step 1 above).
2. Print result via selected method.
3. Toggle between Absorbance and %T mode.
4. Print graph – grayed out if no data are available.
7. Sample number – add a prefix to the sample number and reset the incrementing number to the desired value.
8. Save method – use the left and right arrows to select a folder to store in (Favorites/Methods 1-9), press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing
, or wait.
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p
2: Concent rati on
This makes simple concentration measurements on samples, by measuring the amount of light that has passed through a sample relative to a reference (this can be air). Concentration is obtained by multiplying the measured Absorbance at a specific wavelength by a factor. The factor may be known in advance, or may be calculated by the instrument by measuring a standard of known concentration.
The procedure is as follows:
Step 1
Set wavelength by using keypad numbers or left and right arrows. Press the down arrow key.
Step 2
Select the mode, Factor (user entered) or Standard (factor is calculated from a calibration sample), using the left and right arrows. Press the down arrow key. Step 3 (if Factor is selected) Enter the Factor using the keypad numbers. Range 0.001 to
9999. Use the C button to delete the last digit entered.
Press the down arrow key. Step 3 (if Standard is selected) Enter the concentration using keypad numbers. Range 0.01-
9999. Use the C button to delete the last digit entered.
Press the down arrow key.
Step 4
Units: The user can enter a text string up to 8 characters long. To access a list of pre-defined units press the Options key ::; and then use the left/right arrows (μg/ml, μg/μl, pmol/μl, mg/dl, mmol/l, μmol/l, g/l, mg/l, μg/l, U/l, %, ppm, ppb, conc or none). These units can also be edited once OK is pressed. This screen also allows the number of displayed decimal points (DP) to be selected, from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected (so 98768.2 will display as 98768 even with 1
decimal point selected). Press OK parameters or Cancel
Step 5
To enter the results screen with the selected parameters press
OK OR Cancel the selections and return to the Applications Folder by
pressing Cancel
Step 6 (if using a Factor)
Insert the reference. Press 0A/100% key. This will be used for all subsequent samples until changed.
Step 7
Insert sam
le and press .
.
.
to store the chosen
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Step 6 (if using standard mode) Insert the reference. Press 0A/100% key. This will be used for all subsequent samples until changed.
Press
Run the standard by pressing OR
Press cancel
Step 7
Insert the sample and press The concentration of the sample is displayed. Results shown as
---- indicate the concentration is out of range.
Repeat step 7 for all samples.
Press
Press ::; to display available Options which are described below.
to display the Run Standard screen.
to return to the measure screen.
.
to return to the Applications Folder.
Options (select using key pad numbers)
1. Return to parameters screen (step 1 above).
2. Print result via selected method.
3. Toggles on/off, displaying a graph of wavescan +/- 20 nm from selected wavelength.
4. Return to Run Standard screen.
7. Sample number – add a prefix to the sample number and reset the incrementing number to the desired value.
8. Save method – use the left and right arrows to select a folder to store in (Favorites/Methods 1-9), press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing
, or wait.
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3: Wavescan
An absorption spectrum can be obtained from your instrument, enabling simple identification of peak height and position. The procedure is as follows:
Step 1
Set start wavelength by using keypad numbers or left and right arrows. Press the down arrow key.
Step 2
Set end wavelength by using keypad numbers or left and right arrows. Press the down arrow key.
Step 3
Select the mode, Absorbance or %T, using the left and right
arrows.
Step 4
To enter the measurements screen with the selected parameters
press OK OR Cancel the selections and return to the Applications Folder by
pressing Cancel
Step 5
Insert the reference. Press 0A/100% key. This will be used for all subsequent samples until changed.
Step 6
Insert sample and press
Repeat step 6 for all samples.
Results
A graph of the wavescan is displayed, along with a table of Absorbance/%T at each peak. Use the left and right arrows to move the cursor along the graph. When it reaches a peak the peak height and width of the peak is displayed at the top of the screen. To zoom in on the wavelength scale, use the up arrow. This auto-scales on the Absorbance/%T scale (dependent on the Graph Scale option) and this is retained for subsequent measurements. To zoom out again, use the down arrow.
Press
Press ::; to display available Options which are described next.
.
.
to return to the Applications Folder.
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Options (select using key pad numbers)
1. Return to parameters screen (step 1 above).
2. Print result via selected method.
3. Toggle between Absorbance and %T mode.
4. Displays Peak Detection Parameter Screen. See description below.
5. Manually adds a peak position to the peak table in the results screen at the position set by the cursor. If the cursor is returned to this position the legend “User Defined Peak” is displayed at the top of the scan and this option changes to Delete Peak...
6. Displays Graph Scale Parameter Screen. See description below.
7. Sample number – add a prefix to the sample number and reset the incrementing number to the desired value.
8. Save method – use the left and right arrows to select a folder to store in (Favorites/Methods 1-9), press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing
Peak Detection (Shortcut button 4)
AutoDetect Peaks: Turns on and off the automatic peak
detection. The following options determine how peaks are detected: Minimum peak height: Minimum height the peak has to be above the higher of the two adjacent minima for the peak to be detected Minimum peak width: Minimum width of the peak as determined by the difference in wavelength between the higher of the two adjacent minima and the opposing intersection of that higher minimum level and the peak profile. (See the screen displayed below).
Peak Detect on Zoom: Determines whether peaks are re­assessed and tabulated when the user zooms into a region of the wavescan. If off leaves the peak detection as determined on the un-zoomed display
Sort peaks by…: Determines the sequence that peaks are reported by. Can be wavelength, peak height or peak width.
Draw Peaks: Switches display of peak cursors on and off. These show vertical dashed lines displaying the measured peak height and horizontal dashed lines showing the peak width
Pressing Cancel them.
ignores the selection, pressing accepts
, or wait.
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Add Peak... (Shortcut button 5)
Adds a used defined peak at the current cursor position. The entry is then displayed in inverse coloring to discriminate between user defined peaks and auto-detect peaks. When the cursor is positioned over the user defined peak a legend “User Defined Peak” appears at the top of the graph. The option then changes to Delete Peak to enable the user to remove the peak.
Note: Storing a method at this stage will save these user defined wavelengths, each time method is run Absorbance value at these wavelengths is reported
Graph Scale…
This enables the user to set up a defined graph by defining the limits in either or both of the x and y axes.
Zoom mode:
This sets up the operation of the Zoom keys (up and down arrows). “x & y axes” expands the display around the cursor measurement point, whilst the other options select the absorbance or wavelength axes respectively. With x or y axis limits set to on, zooming out will only be permitted to the set limits.
x/y axis limits:
Setting “x (or y) axis limits” to “On” activates the start and finish points of the desired graph to user defined specific wavelengths and/or absorbance values.
Pressing Cancel them and displays the required graph.
ignores the selection; pressing accepts
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4: Simple Kinetics
Kinetics studies, where the change in absorbance needs to be followed as a function of time at a fixed wavelength, can be readily performed.
Reagent test kits are routinely used for the enzymatic determination of compounds in food, beverage and clinical laboratories by measuring NAD / NADH conversion at 340 nm. The change in absorbance over a specified time period can be used to provide useful information when an appropriate factor, defined in the reagent kit protocol, is applied. Reaction rate and enzyme activity can be calculated if the factor used takes account of the absorbance
difference per unit time, as opposed to the absorbance difference per se.
For this reason, the change in absorbance per minute (ΔA/min), concentration (ΔA/min x factor) and correlation coefficient (calculated from a best fit of the data points) are displayed. They may not be relevant for simple kinetics experiments.
The procedure to define a new method is as follows:
Kinetics Parameters1 Screen Step 1 (Wavelength)
Enter all numerical values using the keypad numbers or the left and right arrows. Use the up and down arrow keys to move between boxes. Step 2 (Delay time) Enter the delay time in seconds before measurements are taken. This can be a maximum of 600 seconds (10 minutes). Step 3 (Duration) Enter the time in minutes over which measurements are taken. This can be a maximum of 60 minutes. Step 4 (Interval) Enter the interval time in seconds between measurements using the left and right arrows. Options are: 5, 10, 20, 30 or 60 seconds.
Step 5
Press Next OR
Press Cancel
Kinetics Parameters 2 Screen Step 6
Select the measurement mode using the left and right arrows. Delta A: change in absorbance over the measurement duration (or selected period). Final A: absorbance at the end of the measurement duration (or selected time). Slope: rate of change of absorbance over the measurement duration or selected period.
to go to the next parameters screen
to return to the Applications Folder.
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Step 7
Units: The user can enter a text string up to 8 characters long. To access a list of pre-defined units press the Options key ::; and then use the left/right arrows (μg/ml, μg/μl, pmol/μl, mg/dl, mmol/l, μmol/l, g/l, mg/l, μg/l, U/l, %, ppm, ppb, conc or none). These units can also be edited once OK is pressed. This screen also allows the number of displayed decimal points (DP) to be selected, from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected (so 98768.2 will display as 98768 even with 1
decimal point selected). Press OK parameters or Cancel
.
to store the chosen
Step 8
Set the Factor by which the result is multiplied to give the amount in the chosen range using the left and right arrows. Range of 0.01 to 9999.
Step 9
Press Next
to enter the Results screen
OR Press Cancel
to return to the Parameters 1 screen.
Results
Insert the reference and press the 0A/100%T key.
Insert the sample and press
to start the run.
Time (min) is displayed at the bottom of the screen, and absorbance data are plotted on the graph as testing proceeds. The table below the graph gives: Absorbance values at T of calculation), T slope, regression parameter (R
(finish of calculation, change in absorbance,
n
2
) of the calculated slope and the
(start
0
result calculated from the selected parameter (dA, final A or slope).
Use the left and right arrows to move the cursor and display the time and absorbance value at measured data points.
Use the up and down arrows to zoom in or out.
Press Cancel
to return to the Applications Folder.
Press ::; to display available Options which are described below.
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Options (select using key pad numbers)
1. Return to parameter 1 screen (step 1 above).
2. Print data on the results screen via selected method.
3. Print all the data.
4. Set the t
position (starting point for the slope and dA
0
calculation) at the current cursor position. Value is retained for subsequent samples.
5. Set the t
position (finishing point for the slope and dA
n
calculation) at the current cursor position. Value is retained for subsequent samples.
6. Toggle the calculated slope line on and off. Note: if any data points enclosed by t
and tn are beyond the
0
range of the instrument (>2.5A or <-0.3A) then this option is grayed out.
7. Sample number – add a prefix to the sample number and reset the incrementing number to the desired value.
8. Save method – use the left and right arrows to select a folder to store in (Favorites/Methods 1-9), press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing
, or wait.
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5: Standard Curve
The construction of a multi-point calibration curve from standards of known concentration to quantify unknown samples is a fundamental use of a spectrophotometer; this instrument has the advantage of being able to store this curve as a method, using up to 9 standards.
To include a zero concentration standard, include this in the number of standards to be entered and enter 0.00 for concentration; use a reagent blank when required to enter the zero standard.
The procedure is as follows:
Step 1
Select the wavelength using the keypad numbers or left and right arrows. Press the down arrow.
Step 2
Enter the number of standard concentration points to be used in the curve (1-9). Press the down arrow.
Step 3
Units: The user can enter a text string up to 8 characters long. To access a list of pre-defined units press the Options key ::; and then use the left/right arrows (μg/ml, μg/μl, pmol/μl, mg/dl, mmol/l, μmol/l, g/l, mg/l, μg/l, U/l, %, ppm, ppb, conc or none). These units can also be edited once OK is pressed. This screen also allows the number of displayed decimal points (DP) to be selected, from 0 to 2 Note that the result will always be fixed to 5 significant figures regardless of how many decimal points are selected (so 98768.2 will display as 98768 even with 1
decimal point selected). Press OK parameters or Cancel
Step 4
Select the type of curve fit using the left and right arrows. Options: straight line regression, a zero regression (this forces the straight line through the origin), interpolated or cubic spline.
Step 5
Select the calibration mode: either Standards (measure prepared standards) or Manual (keypad data entry). Press the down arrow. Step 6 (if standards has been selected in step 5) Select the number of standards to be measured and averaged at each standard concentration point. Can be OFF (1), 2 or 3.
Step 7
Press Next OR
Press Cancel Applications Folder.
to enter the Standards screen
to cancel selections and return to the
.
to store the chosen
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