User Guide
Direct Detect® Spectrometer
C
Made in Germany
The M logo, Millipore, Direct Detect, and Amicon are registered trademarks
of Merck KGaA, Darmstadt, Germany.
All trademarks of third parties are the property of their respective owners.
© 2014 EMD Millipore Corporation, Billerica, MA, U.S.A. All rights reserved.
00000833, Rev. D, 05/14
Notice
The information in this document is subject to change without notice and should not be construed
as a commitment by EMD Millipore Corporation (“Millipore”) or an affiliate. Neither EMD Millipore
Corporation nor any of its affiliates assumes responsibility for any errors that may appear in this
document.
Contents
1 Introduction ............................................................................................................... 1
2 Components Supplied ............................................................................................... 2
3 Symbols Used in this User Guide .............................................................................. 3
4 Product Labeling ........................................................................................................ 3
5 Safety Precautions .................................................................................................... 4
6 Installation ................................................................................................................. 4
6.1 Packaging ........................................................................................................................................................4
6.2 Site Requirements .........................................................................................................................................5
6.3 Installation Configuration ..........................................................................................................................6
6.4 Unpacking and Setting up the Computer ...............................................................................................6
6.5 Unpacking and Setting Up the Direct Detect® Spectrometer ...........................................................6
7 Direct Detect® Spectrometer Parts and Functions .................................................. 8
7.1 General Technical Features .........................................................................................................................8
7.2 Direct Detect® Computer and Software .............................................................................................. 10
8 General Guidelines .................................................................................................. 11
8.1 Buffer components .................................................................................................................................... 11
8.2 Materials Required but Not Supplied ................................................................................................... 11
9 Direct Detect® Assay-free Card and Sample Preparation .....................................12
9.1 Direct Detect® Assay-free Card ............................................................................................................12
9.2 Sample Preparation ................................................................................................................................... 13
10 How to Use the Direct Detect® Spectrometer ....................................................... 14
10.1 Accessing Direct Detect® Software ....................................................................................................... 14
10.2 Loading and Installing the Card ............................................................................................................. 15
10.3 Measurement .............................................................................................................................................. 16
10.4 Removing the Card .................................................................................................................................... 18
11 Switching from Direct Connection to Intranet Connection (Networking) ..........19
11.1 Reconfiguration Requirements ............................................................................................................... 19
11.2 Changing from Direct Connection to Intranet Connection............................................................ 20
11.3 Changing from Intranet Connection to Direct Connection............................................................ 22
11.4 Changing Computer Network Settings Manually ............................................................................. 24
12 Troubleshooting ....................................................................................................... 26
12.1 General Information .................................................................................................................................. 26
12.2 Problems Indicated by Hardware Diagnostics Icon .......................................................................... 27
12.3 Problems Indicated by Instrument Status Light (top of instrument) .......................................... 28
12.4 Problems Indicated by Instrument Validation Icon .......................................................................... 29
12.5 Problem Indicated by Instrument Internal Humidity Icon.............................................................. 29
13 Maintenance ............................................................................................................ 30
13.1 Replacing the IR Source ........................................................................................................................... 30
13.2 Replacing the Desiccant ........................................................................................................................... 33
13.3 Cleaning ........................................................................................................................................................ 35
13.4 Description of Instrument Validation Tests ......................................................................................... 36
13.5 Running the Instrument Validation OQ and PQ Tests ...................................................................... 39
13.6 Storage .......................................................................................................................................................... 39
14 Installation, Operation, and Process Qualification Protocol (Optional) .............. 40
14.1 Installation Qualification Protocol (IQ) ................................................................................................ 40
14.2 Instrument Hardware Diagnostic Protocol..........................................................................................40
14.3 Operation Qualification Protocol (OQ) ................................................................................................. 42
14.4 Performance Qualification Protocol (PQ) ............................................................................................ 42
14.5 Protein Measurement Verification ........................................................................................................ 42
14.6 Direct Detect® System IQ/OQ/PQ Report Summary ......................................................................... 44
15 Specifications........................................................................................................... 45
16 Ordering Information .............................................................................................. 46
17 Technical Assistance ................................................................................................ 47
18 EC Representative .................................................................................................... 47
19 Standard Warranty .................................................................................................. 47
Contents, continued
Direct Detect® Spectrometer User Guide
1
1 Introduction
The Direct Detect® infrared (IR)-based quantification system is an innovative combination of softwarecontrolled instrumentation and EMD Millipore Corporation’s advanced membrane technology, optimized
for biomolecule detection, quantification of proteins, and analysis of lipids/detergents. Biomolecules
are applied directly to a card-based hydrophilic polytetrafluoroethylene (PTFE) membrane that is
transparent in most of the infrared spectral region.
The Direct Detect® system measures amide bonds in protein chains, accurately quantifying an intrinsic
component of every protein without relying on amino acid composition, dye binding properties, or
redox potential. Protein concentrations from 0.25 to 5 mg/mL can accurately be determined from a
minimal sample volume (2 µL) without bio- or immuno-chemical staining. Sample analysis takes only
minutes and in most cases can be performed directly from the buffered or native solution.
Quantification of lipids and detergents requires a user-defined standard curve. Unknown mixtures can
be analyzed by determining the relative IR absorbance.
The system has been validated with proteins and cell lysates representative of those currently in use in
research laboratories.
Direct Detect® Spectrometer User Guide
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2 Components Supplied
The Direct Detect® system includes the following components:
Figure 1. Components supplied
1A
1B
1C
1D
1E
1F
1G
1H
1A Direct Detect® spectrometer
1B Instrument power adapter with 4 power cord/plug configurations (EU, UK, US, Japan)
1C Data cable (crossover, 10Base-T Ethernet with RJ45 connectors)
1D Dell® Netbook computer with Direct Detect® Software installed
1E Computer power adapter with world-wide plug adapter
1F Netbook stand
1G Direct Detect® Assay-free cards (package of 50)
1H Direct Detect® spotting tray
Desiccant pack replacement (1, not shown)
Torx® TX20 screw driver for replacing the IR source (not shown)
Direct Detect® Spectrometer Quick Start Card (not shown)
Direct Detect® Spectrometer User Guide
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3 Symbols Used in this User Guide
The following symbols are used throughout this user guide and/or on product labels, and the user shall
abide by indicated requirements:
Symbol Definition
Y
Warning alerts you to actions that may cause personal injury or pose a
physical threat.
Class I Laser
Product
Class I Laser product, complies with 21 CFR Chapter 1, Subchapter J
i
Read the documentation
h
Catalogue number
f
Serial number
M
Manufacturer
C
CE conformity marking. Reference Declaration of Conformity for specific directives.
Do not discard with common solid waste at end of life. Segregate with other waste
electrical and electronic equipment (WEEE) and send to an appropriate facility for
recycling. For information on recycling electrical and electronic products in the
European Union, please visit www.millipore.com/weee.
Hot Surface
4 Product Labeling
Figure 2. Product label appears on both the back and the bottom of the instrument
Direct Detect® Spectrometer User Guide
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5 Safety Precautions
Review and understand the safety precautions below before installing and operating the Direct Detect®
spectrometer.
Y
WARNING
● To avoid danger of electric shock, do not install the instrument in an area with a high humidity
level. Refer to Site Requirements in section 6.2.
● Do not touch the Ethernet cable or universal power adapter plugs with wet hands.
● To avoid potential shock hazard, choose the correct plug configuration and make sure that the
power adapter is plugged securely into a properly grounded AC power outlet. Make sure that
the connection between the cable and the instrument is secure.
● Always ensure that the power supply input voltage matches the voltage available in your
location.
● Use only the power supplies that were supplied by EMD Millipore Corporation for use with the
Direct Detect® system. Use of other power supplies may damage the system.
● Minimize power draw from other instruments on the same power circuit.
● Do not use with flammable or explosive liquids. Cards that have been spotted with samples in
flammable solvents must be dried prior to insertion into the instrument.
6 Installation
NOTE: For a full installation, operation, and process qualification protocol and checklist, refer to
section14.
6.1 Packaging
Inspect the packaging for damage upon receipt. If there are any signs of damage, contact
the responsible shipping company.
Y
WARNING: Do not put a spectrometer showing signs of damage into operation as this
may result in severe personal injuries and/or property damage.
Keep the original shipping boxes in case you need to return the spectrometer for servicing
or repair.
NOTE: Instruments will not be accepted for repair if not in original packaging. EMD
Millipore Corporation will provide shipping boxes for a charge if original packaging
has been discarded.
Direct Detect® Spectrometer User Guide
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6.2 Site Requirements
The user must provide an installation site that meets the following requirements:
Space requirement Stable and horizontal base which is able to bear the weight of
at least 8 kilograms (kg)
A clearance of at least 25 centimeters (cm) at the
spectrometer rear side
Environment Temperature range: 18–35 °C
Humidity (non-condensing): less than 35% (relative humidity)
Temperature variation: less than 1 °C/hour
Humidity variation: less than 3%/hour
Do not install devices such as large electric motors, heaters,
welding equipment, radio transmitting equipment, units emitting pulsed NMRs, or high-powered lasers in close proximity
to the Direct Detect® spectrometer. These devices can cause a
spectrometer malfunction.
Make sure that these types of devices are not connected to the
same electrical circuit as the Direct Detect® spectrometer.
Vibration Do not locate the Direct Detect® spectrometer near vibration
sources (centrifuges, vortexes, ventilation hoods, air
conditioners, etc.), or in rooms with intense floor vibrations.
Inductive electrical
interference
Do not install the Direct Detect® spectrometer near any
sources of potential inductive electrical interference (e.g.,
pumps, switching motors, microwave ovens, etc.), sources of
high energy pulses, or sources that might cause magnetic or
radio frequency interference.
Power supply Voltage range: 100–240 VAC
Frequency range: 50–60 Hz
Protection class: I
The power supply unit automatically adapts to most power
sources commonly used.
Make sure that the power supply is easily accessible at all
times.
Connect the Direct Detect® spectrometer only to a grounded
power outlet.
Take precautions to ensure uninterrupted power supply if there
is the potential for power supply issues (e.g., brownouts, power
surges, frequent thunderstorms, etc.)
Direct Detect® Spectrometer User Guide
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6.3 Installation Configuration
The Direct Detect® spectrometer is configured to operate as a stand-alone unit
connected to the supplied Netbook computer (PC) using the factory-set standard IP
address (10.10.0.1).
6.4 Unpacking and Setting up the Computer
Unpack the Netbook computer, power adapter, and world-wide plug adapter from
the shipping box. Choose the plug adapter with the correct configuration for local
requirements and attach it to the plug. Plug the low voltage (round) end of the power
adapter into the power port of the PC, but do not plug the computer into a power outlet at
this time.
6.5 Unpacking and Setting Up the Direct Detect® Spectrometer
Remove all components from shipping boxes and lay out on work surface.
6.5.1 Positioning and Connecting Components (Figure 3)
1. Place instrument on bench and put Netbook stand on top of instrument. The
front tab of the stand fits into the slot on top of the instrument (5B in Figure5).
2. Place Netbook computer on stand.
Figure 3. Rear side - Connection ports
3A
3B
3C
3. Turn the instrument/computer assembly so that the rear of the instrument is
facing you (Figure 3).
4. Connect one end of the provided Ethernet cable to the instrument ETH/LAN
port (3A) and the other end to the PC network/LAN port (3B).
Direct Detect® Spectrometer User Guide
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6.5 Unpacking and Setting Up the Direct Detect® Spectrometer, cont.
5. Connect the low voltage (round) end of the instrument power adapter to the
power port of the instrument (3C). Secure the connector with the clamp.
6. Choose the instrument power cord with the correct plug configuration for local
requirements and plug it into the power adapter.
7. Plug both the PC and instrument power cords into a grounded power outlet. A
surge protector is recommended.
Y
WARNING: To avoid personal injuries and/or property damage caused by
electrical power, connect the spectrometer and computer to
a grounded power outlet. Make sure that this power outlet
complies with IEC (International Electrotechnical Commission)
regulations.
8. Turn on the PC.
NOTE: The instrument is powered through the computer and does not have
a separate ON/OFF switch. It should be left on and powered up at all
times.
6.5.2 Initializing the Direct Detect® Spectrometer
1. Refer to the Direct Detect® Software User Guide for instructions on starting up
the software and logging in (section 1.2).
2. During initialization, the laser, IR source, and detector are switched on. The IR
source warms up, the scanner moves forward and backward several times, and
tones once. This phase takes several minutes, and the hardware diagnostics
icon is yellow. After the initialization has been completed successfully, the
hardware diagnostics icon turns to green, but the instrument validation icon
will be yellow (Figure 4). This indicates that the PQ test has expired and needs
to be run. Refer to section 13.5 for instructions on running the PQ test.
Figure 4. Instrument status icons
Instrument validation icon
Instrument internal humidity icon
Hardware diagnostics icon
NOTE: If the status indicator on either the instrument or the software is red or
remains yellow after initialization is complete and the PQ test has been
run, refer to section12, Troubleshooting.
Direct Detect® Spectrometer User Guide
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7 Direct Detect® Spectrometer Parts and Functions
7.1 General Technical Features
7.1.1 The Instrument
The Direct Detect® spectrometer is a small compact FTIR spectrometer designed for
measuring IR absorbance.
Figure 5. Direct Detect® spectrometer
5A
5B
5C
5E
5G
5F
5D
Definition
5A Instrument module (with integrated electronics and optics)
5B Slot for Netbook stand tab
5C Direct Detect® automated card holder
5D Card insertion alignment arrow
5E Status light
5F Sample drying indicator light
5G Card insertion light
7.1.2 The Direct Detect® Automated Card Holder
The automated card holder (5C) accepts only 4-sample Direct Detect® Assay-free cards.
The top of the card holder module contains two LED lights located at the bottom right
corner (5F and 5G as viewed from the top).
The bottom LED light (5G) is green whenever an Assay-free card is inserted properly.
The top LED light (5F) is yellow whenever the drying system heater and fan are active.
Periodically (default setting is 15 minutes) the software will initiate a reference measurement which must be performed without a sample card inserted in the card holder. If a card
is found in the card holder, the software will prompt you to remove it.
Direct Detect® Spectrometer User Guide
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7.1.3 Connecting Ports and LEDs
The connecting ports (e.g., power, Ethernet) and the system diagnostic LEDs are on the
back of the instrument.
Figure 6. Direct Detect® Spectrometer rear side - Connecting ports and LEDs
6A
6B 6C 6E 6F 6G 6H 6I 6J 6K
6D
Type Definition
6A
USB port Not available to user. For instrument service use only.
6B
ETH/LAN port The ETH/LAN port connects the spectrometer to the Netbook
computer, using the supplied data cable (crossover, 10Base-T with
RJ45 ports).
6C TKD LED The green TKD LED indicates that the interferometer mirror is
within the data acquisition range. When data acquisition is
occurring, the light is bright green. The abbreviation TKD means
Take Data.
6D ACC LED The yellow ACC LED indicates that the network is accessing the
optics. During transition of measuring data the LED is light yellow.
The abbreviation ACC means Access.
6E DIR/ERR LED In normal operation mode the DIR/ERR LED flashes pale red and
indicates scanner movement. If the LED lights permanently bright
red, there is an interferometer problem, e.g., missing laser signal.
When this LED is permanently red no data acquisition is possible.
The abbreviation DIR means Direction and ERR means Error.
6F External
accessory port
Not available to user. For instrument service use only.
6G TAP port Not available to user. For instrument service use only.
7.1 Direct Detect® Spectrometer General Technical Features, continued
Direct Detect® Spectrometer User Guide
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Type Definition
6H CFG switch CFG is a four-fold DIP switch:
#1 ON and #2 OFF: DHCP mode - use last DHCP address
#1 OFF and #2 ON: use ON-IP address 10.10.0.1
#3: stay in boot loader, do not load the application program
#4: reserved
The abbreviation CFG means Configuration.
6I SBY/RES button Pressing this button for approximately 2 seconds, activates/
deactivates the stand-by mode. Pressing this button for more
than 6 seconds resets the spectrometer to the original state. The
abbreviation SBY means Standby and RES means Reset. Do not use
unless instructed to do so by service personnel.
6J POWER port This port connects the spectrometer to the power supply via the
supplied power adapter and power cord. The POWER port is for
extra low-voltage connectors. Use only power cords/supplies that
have been supplied by EMD Millipore Corporation.
6K Kensington lock
slot
The lock slot is an anti-theft lock for mobile hardware. Lock and
cable are not supplied.
7.2 Direct Detect® Computer and Software
The Direct Detect® spectrometer is designed to operate with the Direct Detect® Software
that is pre-installed on the Netbook computer. For computer specifications, refer to
section 15.
Direct Detect® Software may be updated periodically. For information on the most up-todate software, go to www.millipore.com/directdetect. Register your instrument in order to
receive notification about relevant software upgrades.
WARNING: Do not install third-party software (screen savers, etc.), as this may impact the
performance of the Direct Detect® system.
7.1 Direct Detect® Spectrometer General Technical Features, continued
Direct Detect® Spectrometer User Guide
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8 General Guidelines
The Direct Detect® spectrometer is recommended for use with protein samples ranging from 0.25 to
5mg/mL. Although the instrument can measure protein concentrations from 0.1 mg/mL to around
30mg/mL, the most accurate results are achieved within the recommended concentration range. The
instrument is also capable of detecting and analyzing lipids and detergents. Analysis is performed with
a user-defined calibration method. Samples to be analyzed are introduced to the system using the
Assay-free card which contains a novel IR-compatible membrane. The recommended sample volume is
2 µL per spot position.
8.1 Buffer components
The Direct Detect® system is compatible with most commonly used buffers, detergents, and
reducing agents, however, use of buffers which contain amides or amine functional groups
may affect quantification.
Buffers containing more than 5% glycerol, SDS, and/or Tween® 20 surfactant should be
avoided due to the excessive time required to achieve dryness.
The instrument comes with preloaded protein calibration files, prepared using National
Institute of Standards & Technology (NIST) bovine serum albumin (BSA) in phosphatebuffered saline (PBS). However, preparation of a fresh calibration method with the
buffer used for protein preparation is recommended. The Direct Detect® spectrometer is
compatible with most solvents used in lipid research.
8.2 Materials Required but Not Supplied
● Protein standards (Bovine Serum Albumin - National Institute of Standards &
Technology SRM 927d (optional))
● Protein sample(s)
● Lipid/detergent standards
● Lipid/detergent sample(s)
● Fixed-volume precision pipette capable of precisely dispensing 2 µL, and pipette tips
NOTE: To minimize pipetting error, we recommend the use of a fixed-volume precision
pipette capable of dispensing 2 µL, for applying samples to the Assay-free card.
Adjustable pipettes are not recommended.
● Water, PBS, or other buffer
● Chloroform or other lipid compatible solvent
● Printer (optional)
Direct Detect® Spectrometer User Guide
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9 Direct Detect® Assay-free Card and Sample Preparation
9.1 Direct Detect® Assay-free Card
All concentration measurements are performed using the Assay-free card.
Figure 7. Direct Detect® Assay-free card
The card contains four hydrophilic polytetrafluoroethylene (PTFE) membrane positions,
sized for easy sample application and surrounded by a hydrophobic ring that retains the
sample to be analyzed within the IR beam. A 2 µL sample volume is recommended in order
to assure proper sample application, drying, and measurement.
By design, the positions on the card are numbered from 1 to 4, starting from the bottom.
Position 1, by software default, is reserved for the “blank” measurement. The “blank” is the
buffer/solvent used to resuspend the sample, but contains no protein or lipid/detergent.
One or more of the remaining card positions are used for the protein and/or lipid/detergent
samples.
The Assay-free card can be inserted into the system and analyzed immediately after
preparation. The Direct Detect® spectrometer has a built-in drying system that
automatically dries the sample to the point where it gives an accurate reading. The first
sample position typically takes the longest period of time to dry, with subsequent positions
taking less time. Including drying time, the total measurement time for a card loaded with
2 µL samples does not exceed 5 minutes.
The Direct Detect® Assay-free card is intended for one-time protein and/or lipid/detergent
spotting. However, already prepared cards can be stored and read again.
Direct Detect® Spectrometer User Guide
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9.2 Sample Preparation
In general, analysis of protein samples within the concentration range of 0.25–5 mg/mL
can be performed directly from the buffered solution. Buffers containing more than 5%
glycerol, SDS, and/or Tween® 20 surfactant should be avoided due to the excessive time
required to achieve dryness.
For analysis of samples with protein content above 5 mg/mL, single step dilution is
recommended.
Concentration determination of samples below 0.25 mg/mL can be achieved after a simple
spin concentration step using a centrifugal filter (e.g., Amicon® Ultra-0.5 device).
Use of centrifugal filters is also recommended for buffer exchange when the original buffer
contains a high concentration of components with amide- or amine-containing functional
groups. Figure 8 is an example of a diafiltration protocol that concentrates the sample and
reduces the Tris buffer concentration.
500 µL of
0.1 mg/mL
protein in
50 mM Tris
50 µL of
1 mg/mL
protein in
50 mM Tris
50 mM
Tris
50 µL of
1 mg/mL
protein in
PBS with 9.5
mM Tris
PBS with
9.5 mM Tris
Add 450 µL of
1X PBS or
exchange buffer
500 µL of
0.1 mg/mL
protein in
PBS with 9.5
mM Tris
Spin to
concentrate
Spin to
concentrate
Figure 8. Buffer exchange and concentration using an Amicon® Ultra-0.5 device
In general, lipid sample preparation involves solvent extraction. Neutral lipids may be
extracted by relatively nonpolar solvents such as hexane, chloroform, or toluene while
membrane-associated lipids require polar solvents like methanol or ethanol.
Lipids dissolved in organic solvents may be applied directly onto the Assay-free card.
Special care in the application of highly volatile solvents (chloroform, hexane, etc.) is
recommended in order to contain the samples within the hydrophobic embossing.
Analysis of detergents can be performed directly from buffer. Due to the excessive time
required to achieve dryness, samples containing more than 5% of the analyzed detergent
should be avoided.
The limits of lipid and detergent detection and quantification should be established
experimentally for each compound analyzed.
Direct Detect® Spectrometer User Guide
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10 How to Use the Direct Detect® Spectrometer
The instrument is controlled and operated by Direct Detect® Software installed on the Netbook
computer. Samples are introduced to the instrument module by inserting the Direct Detect® Assay-free
card into the card holder (Figure 11). An alignment notch on one corner of the card ensures correct
card insertion into the card holder.
10.1 Accessing Direct Detect® Software
The Direct Detect® Software can be accessed through the icon on the Netbook desktop.
Figure 9. Direct Detect® Software desktop icon
Each instrument user can be assigned a unique name and optional password (Figure 10).
NOTE: If 21CFR11 mode is enabled, a password is mandatory. Refer to section 1.3 in the
Direct Detect® Software User Guide.
The measurements performed by each user are stored and recalled based upon the user
name.
Figure 10. Direct Detect® login screen
For detailed instructions on starting up the software, refer to section 1.2 of the Direct
Detect® Software User Guide.
NOTE: A seven minute warm-up cycle is required after initial power-up and software
activation. The software will wait for seven minutes before allowing a sample
measurement to begin.
Direct Detect® Spectrometer User Guide
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10.2 Loading and Installing the Card
1. Place the Assay-free card(s) on the spotting tray or other clean surface. Select an
appropriate buffer/solvent blank, preferably the same solution in which the protein
or lipid/detergent has been prepared. If that buffer/solvent is not fully characterized,
select the closest buffer/solvent possible. Pipette 2 µL of the buffer/solvent solution to
the default blank position on the card (position1).
2. Pipette 2 µL of sample to sample positions 2 through 4 as needed. Unused positions can
be loaded and read at a later time as long as the buffer/solvent blank is appropriate for
them. Card positions can be labeled if desired.
NOTE: Cards spotted with samples in flammable solvents must be dried at ambient
conditions prior to inserting the Assay-free card into the instrument. However,
for nonflammable samples, the Direct Detect® spectrometer has a built-in
drying system that can automatically dry the sample to the point where it will
give an accurate reading.
3. Insert the card vertically into the slot in the top of the card holder module with the
card facing left and the alignment notch in the corner of the card oriented towards the
center of the instrument.
Figure 11. Inserting sample card
The arrow on the bottom right side of the card should be aligned with the arrow on
the Direct Detect® card holder module (5D in Figure 5). Gently release the card and the
instrument will accept it for analysis.
When the card is inserted properly the instrument will respond by jogging the card
holder up and down briefly. The bottom green LED on Direct Detect® card holder
module will light and the system will produce a brief tone confirming correct
placement of the Assay-free card.
NOTE: The software will not allow sample measurement if the card is not inserted
properly.
Direct Detect® Spectrometer User Guide
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10.3 Measurement
For detailed instructions on setting up a sample measurement, refer to section 2.2 of the
Direct Detect® Software User Guide.
10.3.1 General Measurement Procedure
1. Complete the sample measurement setup (Figure 12). Check the Dry sample
card box to allow the instrument to dry the sample.
Figure 12. Sample information
NOTE: The Direct Detect® system comes with three preloaded protein
calibration methods. These pre-existing methods may be used for
all future measurements, however, for accurate quantification, we
recommend preparation of a fresh calibration method for each buffer
used in sample preparation (refer to section 4.2 in the Direct Detect®
Software User Guide). Lipids and detergents can be analyzed using a
Relative Absorbance mode.
2. Click on the Measure Card button (Figure 13). The sample measurement takes
about a minute and the measurement progress is shown in the lower status bar.
Figure 13. Measure sample
Direct Detect® Spectrometer User Guide
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The result from each measured sample position is displayed in the Last Sample
Result tab (Figure 14) of the main Direct Detect® screen.
Figure 14. Last sample results screen
3. For reporting options, refer to sections 2.4–2.7 of the Direct Detect® Software
User Guide.
NOTE: Periodically the software will initiate a reference measurement which
must be performed without a sample card inserted in the card holder.
If a card is found in the card holder, the software will prompt you to
remove it.
10.3.2 Calibration
The Calibration Method Wizard (Options menu, Generate Calib. Curve) is used to
create or modify calibration method files that will be used to perform the analysis
of the Assay-free cards.
Figure 15. Calibration mode
To create a calibration method you need previously collected spectra from a series
of known protein or lipid/detergent concentrations. To compile these files, select
Acquire Calibration Spectra; Protein Region or Lipids Region from the Options
menu and follow the instructions for collecting calibration spectra in section 4.2 of
the Direct Detect® Software User Guide.
10.3 Measurement, continued
Direct Detect® Spectrometer User Guide
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Section 4.2 of the Direct Detect® Software User Guide details how to use the
Calibration Method Wizard to create or modify a calibration method file.
Figure 16. Calibration Method start page
10.4 Removing the Card
After all 4 positions on the card have been read, the instrument will sound a tone. The card
will rise to the initial insertion position and can be removed.
CAUTION: Do not remove card during the reading. To stop a reading in progress, click on
the Abort button.
Figure 17. Abort button
When a measurement has been aborted, the progress bar will display Measurement
Aborted. After a short period of time, the instrument will return to Measure Card status.
10.3 Measurement, continued
Direct Detect® Spectrometer User Guide
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11 Switching from Direct Connection to Intranet
Connection (Networking)
During initial setup, the Direct Detect® instrument is connected directly to the computer running
the Direct Detect® software using the crossover 10Base-T cable with RJ45 connectors provided with
the instrument (direct connection). After verification of instrument and software functionality, the
instrument can be configured to run directly from your company’s intranet (intranet connection), if
desired.
11.1 Reconfiguration Requirements
1. Confirm that the system initializes and run the PQ test (refer to section 13.5). If a full
Installation, Operation, and Process Qualification Protocol is required, perform all tests
prior to changing the connections (refer to section 14).
2. The IP Address Change Utility is located in C:\directdetect\IPAddressChanger. This
utility assists in the transition between direct connection to the computer and
connection via the intranet and vice versa. Two standard RJ45Cat5e Ethernet cables
(not provided) are required to switch to an intranet connection.
When the IP Address Change Utility starts, the screen lists the requirements necessary
to perform the change in operating mode (Figure 18):
● The Direct Detect® software is not running on the computer.
● The instrument is powered on and connected, either directly to the instrument or to
the intranet.
● The software is able to locate the Direct Detect® installation folder and files
required to reconfigure the instrument.
Figure 18. IP address change utility requirements
3. Select the Next button and the software will check the validity of each requirement.
Direct Detect® Spectrometer User Guide
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11.1 Reconfiguration Requirements, continued
4. If a requirement is found to be valid, a appears next to the requirement in the list.
If a requirement is not valid then an will appear next to the requirement. A message
indicating the cause of the failure will appear.
You can continue to the next step in the utility only if all requirements are valid.
Figure 19. Status of requirements
11.2 Changing from Direct Connection to Intranet Connection
IMPORTANT: This section assumes that the Direct Detect® instrument is currently
connected directly to the computer using the supplied crossover 10Base-T cable with RJ45
connectors.
1. If the requirements are valid (all check marks are green) select the Next button to
proceed to the IP Settings selection screen of the utility.
2. Select the second option, I want to connect the instrument to the network using
IPAddress.
Figure 20. Selecting network IP address
3. An IP address URL field will become active to the right of this option. Enter the fixed IP
address issued by your IT department. The URL must be fixed for this instrument.
4. Select the Next button to proceed.
Direct Detect® Spectrometer User Guide
21
5. The software will now make the necessary modifications to the Direct Detect® software
and instrument, as well as the host computer. If the modifications are successful you
will be informed that your instrument settings have been updated.
Figure 21. Successful reconfiguration to intranet connection
6. Unplug the instrument from the power source, then change the switches in the red
4-position DIP switch (6H in Figure6) on the rear of the instrument so that all four are
in the DOWN position (refer to Figure 21).
7. Select the Next button to proceed.
8. Run one of the standard RJ45 Cat5e Ethernet cables between the ETH/LAN port on the
rear of the instrument (6B in Figure 6), and your intranet hub. Run the other standard
RJ45 Cat5e Ethernet cable between the PC network/LAN port (3B in figure 3) of the
Netbook computer and your intranet hub. The supplied crossover 10Base-T cable with
RJ45 connectors is not required for this configuration.
9. Reconnect power to the instrument.
10. Wait for the instrument to initialize and then select the Test Connection button on the
utility screen.
Figure 22. Testing the connection
11.2 Changing from Direct Connection to Intranet Connection, continued
Direct Detect® Spectrometer User Guide
22
11.2 Changing from Direct Connection to Intranet Connection, continued
11. If the connection was successful, the instrument will produce a tone and a message
box will appear to inform you that the connection was successful. If an error message
appears, check with your IT department or contact Technical Service (section 17).
12. Select the Finish button to close the software. You should now be able to perform
measurements using the Direct Detect® instrument connected to your intranet.
11.3 Changing from Intranet Connection to Direct Connection
IMPORTANT: This section assumes that the Direct Detect® instrument and Netbook
computer are currently connected to your intranet using two standard RJ45 Cat5e
Ethernet cables.
1. Run the IP Address Change Utility (section 11.1) and confirm that the requirements
are all valid (green check marks).
2. Select the Next button to proceed to the IP Settings screen of the utility and select the
first option, I want to connect the instrument directly to the computer.
Figure 23. Selecting direct connection
3. Select the Next button to proceed.
4. The software will now make the necessary modifications to the Direct Detect® software
and instrument, as well as the host computer. If the modifications are successful you
will be informed that your instrument settings have been updated (Figure 24).
Direct Detect® Spectrometer User Guide
23
Figure 24. Successful reconfiguration to direct connection
5. Unplug the instrument from the power source, then change the switches in the red
4-position DIP switch (6H in Figure6) on the rear of the instrument so that the second
switch from the left is in the UP position, and all other switches are in the DOWN
position (refer to Figure 24).
6. Select the Next button to proceed.
7. Run the crossover 10Base-T cable with RJ45 connectors supplied with your instrument
between the ETH/LAN port on the rear of the instrument (6B in Figure 6) and the
network/LAN port on the computer (3B in Figure 3).
8. Reconnect power to the instrument.
9. Wait for the instrument to initialize and then select the Test Connection button on the
utility screen (Figure 25).
Figure 25. Testing the connection
10. If the connection was successful, the instrument will produce a tone and a message
box will appear to inform you that the connection was successful. If an error message
appears, check with your IT department or contact Technical Service (section 17).
11. Select the Finish button to close the software. You should now be able to perform
measurements using the Direct Detect® instrument connected directly to the computer.
11.3 Changing from Intranet Connection to Direct Connection, continued
Direct Detect® Spectrometer User Guide
24
11.4 Changing Computer Network Settings Manually
Typically the IP Address Changer software makes the appropriate modifications
automatically, however, in some cases you may need to do it manually.
1. To access the settings in the Windows® 7 operating
system, enter TCPIP in the search field. Select the
View network connections option in the control
panel section.
Figure 26. Accessing network connections in the Windows® 7 operating system
2. Select your network connection, and then select option Change settings of this
connection.
3. This opens the Local Area Connection Properties dialog. Select the connection
Internet Protocol Version 4(TCP/IPv4) and then select the Properties button to view
this connection’s properties.
Figure 27. Accessing the connection’s properties
Direct Detect® Spectrometer User Guide
25
4. Depending on the desired configuration, choose one of the following options:
For a direct connection between the Direct Detect® instrument and the computer,
enter the custom IP address 10.10.0.100 and subnet mask setting 255.255.255.0
(Figure28).
IMPORTANT: Do not change the settings on your computer until after you have run the
IP Address Change Utility to change the IP address on the instrument and the Direct
Detect® software (section 11.2).
Figure 28. IP address and subnet mask settings for direct connection
OR
For a connection between the Direct Detect® instrument and the computer via
your intranet, consult with your IT department. Typically the settings are set to obtain
the IP address automatically. However, some IT departments may issue fixed addresses.
Figure 29. IP address and subnet mask settings
IMPORTANT: Do not change the settings on your computer until after you have run the
IP Address Change Utility to change the IP address on the instrument and the Direct
Detect® software (section 11.3).
11.4 Changing Computer Network Settings Manually, continued
Direct Detect® Spectrometer User Guide
26
12 Troubleshooting
12.1 General Information
The status of the Direct Detect® spectrometer is indicated by both the instrument
status light (located on the top right side of the instrument under the Netbook tray (5E
in Figure5)) and the hardware diagnostics icon (located in the upper right corner of
the Direct Detect® Software user interface (Figure 30)). The status light and hardware
diagnostics icon colors are typically the same at any given time, and indicate the
following:
Green Instrument is in proper operating condition (flashing green on the instrument
indicates stand-by mode)
Yellow Instrument is initializing or indicating a warning (e.g., IR source has not yet
reached its operating temperature, the validity period of the instrument has
expired, the air humidity inside the spectrometer is too high)
Red Instrument has malfunctioned, or has a defective component, or an OQ/PQ test
has expired or failed
Figure 30. Direct Detect® Software user interface
Instrument internal
humidity icon
Instrument validation icon
and OQ/PQ initiator
Hardware diagnostics icon
Direct Detect® Spectrometer User Guide
27
Click on the hardware diagnostics icon and the hardware diagnostics dialog opens.
Figure 31. Hardware diagnostics dialog - Instrument Status
More information on hardware diagnostics is available in section 5.1 of the Direct Detect®
Software User Guide. If an issue cannot be resolved with the help of this diagnostics
section and/or the following troubleshooting tables, refer to section17 for information on
how to contact Technical Service.
12.2 Problems Indicated by Hardware Diagnostics Icon
Symptom Cause Corrective Action
Hardware
diagnostics icon
is yellow
Spectrometer is still in the
initialization phase.
Wait until spectrometer initialization is completed
(several minutes).
IR source has not yet
reached its operating
temperature.
The warm-up takes about 7 minutes. As soon as the
IR source has reached its operating temperature, the
hardware diagnostics icon will automatically turn
green. Wait until the warm-up is completed.
IR source is nearing end of
operational lifetime.
To confirm IR source issue, click on the hardware
diagnostics icon (Figure 30) to open the hardware
diagnostics dialog (Figure 31). If IR source is indicated,
refer to section 13.1 for instructions on how to replace
it.
Instrument internal
humidity is above tolerance
(also indicated by the
yellow internal humidity
icon, see section 12.5).
Wait 30 minutes to 24 hours for desiccant to remove
the humidity. If after 24 hours the icon is still yellow,
refer to section 13.2, Replacing the Desiccant.
12.1 General Information, continued
Direct Detect® Spectrometer User Guide
28
Symptom Cause Corrective Action
Hardware
diagnostics icon
is red
Instrument test has failed. To determine which instrument component is causing
the test failure, click on the hardware diagnostics icon
(Figure 30) to open the hardware diagnostics dialog
(Figure 31). If the laser, interferometer, electronics,
automation, or detector is defective, contact Technical
Service. If the IR source is defective, refer to section
13.1 for instructions on how to replace it.
A spectrometer component
is defective.
To find out which component is defective, click on
the hardware diagnostics icon (Figure 30) to open the
hardware diagnostics dialog (Figure 31). If the laser,
interferometer, electronics, automation, or detector is
defective, contact Technical Service. If the IR source is
defective, refer to section 13.1 for instructions on how
to replace it.
12.3 Problems Indicated by Instrument Status Light (top of instrument)
Symptom Cause Corrective Action
Spectrometer
status light does
not light
Spectrometer is not
connected properly to the
power supply.
Check the power cord connection at the power outlet
and the power port on the back of the spectrometer
(Figure 3).
No voltage is applied. Confirm proper voltage at the power outlet.
Short circuit in the adapter
or power cable.
A short circuit is accompanied by a “ticking” sound in
the power adapter. Unplug the power cord immediately
and contact Technical Service.
Spectrometer
status light is
yellow
Spectrometer is still in the
initialization phase.
Wait until spectrometer initialization is completed
(several minutes).
IR source has not yet
reached its operating
temperature.
The warm-up takes about 7 minutes. As soon as the
IR source has reached its operating temperature the
status light will automatically turn green. Wait until
the warm-up is completed.
Spectrometer
status light is red
Instrument test has failed. To determine which instrument component is causing
the test failure, click on the hardware diagnostics icon
(Figure 30) to open the hardware diagnostics dialog
(Figure 31). If the laser, interferometer, electronics,
automation, or detector is defective, contact Technical
Service. If the IR source is defective, refer to section
13.1 for instructions on how to replace it.
A spectrometer component
is defective.
To find out which component is defective, click on
the hardware diagnostics icon (Figure 30) to open the
hardware diagnostics dialog (Figure 31). If the laser,
interferometer, electronics, automation, or detector is
defective, contact Technical Service. If the IR source is
defective, refer to section 13.1 for instructions on how
to replace it.
12.2 Problems Indicated by Hardware Diagnostics Icon, continued
Direct Detect® Spectrometer User Guide
29
12.4 Problems Indicated by Instrument Validation Icon
Instrument validation problems are indicated by the Instrument Validation Icon (Figure 30).
Symptom Cause Corrective Action
Instrument
validation icon
is yellow
An instrument validation
test has expired.
To find out which instrument validation test has
expired, click on the instrument validation icon to
open the instrument validation dialog. Refer to section
5.2 of the Direct Detect® Software User Guide for
details on instrument validation.
Instrument
validation icon
is red
An instrument validation
test has failed.
To find out which instrument validation test has failed,
click on the instrument validation icon to open the
instrument validation dialog. Refer to section 5.2 of
the Direct Detect® Software User Guide for details on
instrument validation.
12.5 Problem Indicated by Instrument Internal Humidity Icon
Symptom Cause Corrective Action
Internal humidity
icon is yellow
Instrument internal
humidity is above
tolerance.
Click on the internal humidity icon to see what the
current humidity level is. If the instrument has been
on for more than 60minutes, and the humidity
warning is still displayed, replace the desiccant packets
in the rear compartment of the instrument. Refer to
section 13.2.
Direct Detect® Spectrometer User Guide
30
13 Maintenance
The Direct Detect® spectrometer is a low-maintenance instrument and the operator can replace
components with a limited service life. Replacing the IR source and desiccant are described in this
section.
Y
WARNING
Perform only the maintenance procedures described in this manual and observe the relevant safety
precautions. Failure to do so may cause property damage or personal injury.
Maintenance or repair procedures not described in this manual should be performed only by an
EMD Millipore Corporation service engineer. Refer to the Technical Assistance section for contact
information.
13.1 Replacing the IR Source
13.1.1 IR Source Status Indicator
The Direct Detect® spectrometer is equipped with a highly-efficient, low-power,
air-cooled infrared source with an average life span of 44,000 hours. As the end
of this life span is approached, the hardware diagnostics icon turns yellow. When
you click on the hardware diagnostics icon (Figure 30), the hardware diagnostics
dialog appears and displays the message End of lifetime is nearly reached
(Figure31). Samples can still be measured, but performance and reliability may
be compromised and the IR source (cat. no. DDSP00010-LA) should be replaced as
soon as possible.
If the source is defective, the hardware diagnostics icon turns red. When you click
on this icon, the hardware diagnostics dialog appears and displays the message No
source is connected. Samples can no longer be measured, and the IR source must
be replaced.
13.1.2 Replacing the IR Source
WARNING: During spectrometer operation the light source becomes very
hot. To avoid burns, wait until the light source has cooled down
before replacing it.
1. Unplug the instrument power plug.
2. On the back of the instrument, loosen the 4 cover plate Torx® screws with the
Torx®TX20 screw driver supplied (Figure32).
Direct Detect® Spectrometer User Guide
31
Figure 32. Removing the cover plate
Torx® screws
3. Take off the cover plate.
Figure 33. Removing the IR source
IR source
Source plug
IR source
Torx® screws
Alignment
groove
4. Loosen the two mounting screws on the IR source (Figure33) with the Torx®
screw driver.
5. Disconnect the green source plug (Figure33).
6. Pull the source out from the spectrometer housing.
13.1 Replacing the IR Source, continued
Direct Detect® Spectrometer User Guide
32
Figure 34. IR source removed from instrument
Alignment
groove
Alignment
pin
7. Insert the new IR source into the spectrometer housing. Make sure that the
alignment pin fits into the alignment groove (Figure 34).
8. Secure the source by fastening the two Torx® screws (Figure35) .
9. Plug the green source plug into the SRC female connector.
Figure 35. Replacing IR source Torx® screws
10. Attach the cover plate and fasten the 4 Torx® screws (Figure 32). Plug the
power plug into the power outlet again.
13.1 Replacing the IR Source, continued
Direct Detect® Spectrometer User Guide
33
After replacing the IR source, perform the Performance Qualification and
Operational Qualification instrument tests as follows:
11. Click on the instrument validation button .
12. When the instrument validation dialog appears, click on the Performance
Qualification Run Test button (1 in Figure36). The test takes about 5 minutes.
Figure 36. Instrument validation dialog
2
1
13. After the test has finished, run the Operational Qualification test by clicking on
the Operational Qualification Run Test button (2).
14. If the instrument passes the test, the hardware diagnostics icon turns green.
If the instrument fails, refer to section 5.1 in the Direct Detect® Software User
Guide.
13.2 Replacing the Desiccant
Excessive humidity inside the spectrometer may cause H2O bands in the spectrum. To
reduce H2O bands to a minimum, the air inside the Direct Detect® instrument module is
convectively dried with a desiccant enclosed in a small, removable, multichambered pack.
If the desiccant is saturated and can no longer absorb humidity, it must be replaced.
Both the internal humidity icon (Figure 37) and instrument status light (on top of the
instrument, 5E in Figure 5) will turn yellow. When you click on the internal humidity icon,
a warning message with the current humidity level is displayed (Figure 37).
13.1 Replacing the IR Source, continued
Direct Detect® Spectrometer User Guide
34
Internal humidity icon
Figure 37. Instrument internal humidity warning
Measuring is still possible if the status light is yellow, but the desiccant should be replaced
as soon as possible.
Replace the desiccant as follows:
1. Unplug the power plug.
2. The desiccant is accessible from the rear side of the spectrometer. Loosen the 4Torx®
screws (Figure 32) with the Torx® screw driver supplied and take off the cover plate.
Figure 38. Replacing the desiccant
Desiccant
pack
3. Pull out the used desiccant pack. A pair of tweezers may be helpful.
13.2 Replacing the Desiccant, continued
Direct Detect® Spectrometer User Guide
35
13.2 Replacing the Desiccant, continued
4. Remove the external packaging from the replacement desiccant pack.
5. Insert the desiccant pack properly into the spectrometer as shown in Figure38.
Y
CAUTION: Do not cut or tear the pack into single pieces or ingest the desiccant.
6. Attach the cover plate and fasten the 4 Torx® screws again (Figure32).
7. Plug the power plug into the power outlet again. After the new desiccant removes the
moisture from the chamber (approximately 30 minutes), the instrument status light
and hardware diagnostics icon will turn back to green.
13.3 Cleaning
The outer spectrometer surface can be cleaned with a damp or dry cloth. Do not use
detergents with organic solvents, acids, or bases!
Y
CAUTION: Do not try to clean the interior of the spectrometer as this may lead to
serious spectrometer damage.
Direct Detect® Spectrometer User Guide
36
13.4 Description of Instrument Validation Tests
13.4.1 Performance Qualification Test (PQ)
The Performance Qualification test evaluates the spectroscopic performance and
confirms proper function of the IR spectrometer. To ensure that the instrument is
working properly, it is recommended that the PQ test be run on a weekly basis.
The factory default interval for the PQ test is one week. One week from the last PQ
test, the software will alert you to run the instrument PQ test again. You can use
the instrument with an expired PQ test, but the software will query you each time
a measurement is initiated, asking if you wish to proceed under the expired PQ
condition.
The PQ test protocol consists of the following three tests:
●Signal-to-noise test: The signal-to-noise ratio determines the sensitivity of
the spectrometer by calculating the average signal-to-noise ratio of ten 100%
spectra. A 100% spectrum is the ratio of two successively acquired singlechannel spectra with no sample in the sample compartment. The ratio of these
two single-channel spectra is used to generate a transmission spectrum.
●100% Line Test: This test verifies that the 100% line does not shift over time.
It measures the maximum deviation of a 100% line within a larger frequency
range. The average of ten measurements must not exceed the predefined limit.
●Wavenumber Accuracy Test: - This test ensures that the frequency calibration
of the instrument is within the defined limits using an internal polystyrene
standard sample.
Direct Detect® Spectrometer User Guide
37
Figure 39 shows a successful PQ test report.
OVP - PQ Test Protocol
Company :
Operator:
Instrument Ty pe:
Optics Configuration:
Accessory:
Instrument Serial Number:
Instrument Fir mwar e Ver sion:
OPUS/DB Version:
Overall Tes t Result
Test expires:
Test Date/Time:
Test Spectra Path:
Date of last PQ Ref erenc e Measurement
Comment:
MILLIPORE
Default
Alpha Sample Compartment RT-DLaTGS ZnSe
Fibre 1 w ith: MIR, KBr, RT- DLATGS [Internal Pos.1]
Millipore Dir ect-Det ec t
M3
1.320_MDD_01 Oct 27 2011
OPUS 7.0 Build: 7, 0, 122 / DB: 7.0.105.1106 for OPUS 7.0
PA SSED
12/25/2011, 3:12:41 PM (GMT-5)
12/15/2011, 3:12:41 PM (GMT-5)
C:\Dire ct Detect\V alidation\Data\20111215\151241
Signal to Noise Te st
Minim um Signal to Noise L imit (are a 1): 1000 Signal to Noise m eas ured: 3458
100% Line T es t
Maxim um 100% Line Deviation: 2.0 M eas ured 100% Line Deviation: 0.29
Wavenum ber Accur acy Te st - Polystyr ene (35.5 Deg. C, Pe ak is T. cor r.)
Sample Mater ial:
Specifie d Peak:
Measured Peak:
Cor re cted Pe ak:
1601.45 cm -1
1601.13 cm -1
1601.25 cm -1
Polys tyren e
Maxim um Deviation:
Me asu re d Deviation :
0.50 cm -1
0.20 cm -1
Overall Test Result = PASSED
Figure 39. PQ test report
13.4.2 Operational Qualification Test (OQ)
The Operational Qualification test checks instrument performance and compares
this performance to the instrument specifications. The OQ test is performed during
final testing before an instrument is delivered to the customer. It should also be
performed after each major repair, after replacement of optical components which
may influence the instrument performance, and after maintenance has taken place.
The factory default interval for the OQ test is 12 months. Twelve months from the
last OQ test, the software will alert you to run the instrument OQ test again. You
can use the instrument with an expired OQ test, but the software will query you
each time a measurement is initiated, asking if you would like to proceed under the
expired OQ condition.
The OQ test protocol consists of the following four tests:
●Wavenumber Resolution Test: This test verifies the achievable spectrometer
resolution by measuring a single-channel (single-beam) spectrum of the water
vapor available inside the spectrometer, using the maximum available resolution.
13.4 Description of Instrument Validation Tests, continued
Direct Detect® Spectrometer User Guide
38
●Signal Sensitivity Test: This test is the same as the PQ signal-to-noise test,
except that the spectra are measured in a very small region (2,200–2,100 cm-1).
●Energy Distribution Test: This test verifies the achievable frequency range and
the energy at the maximum position of the single-channel spectrum. Absolute
values of energy are measured at different wavenumber positions. These values
are divided by the maximum value achieved, and compared to acceptance values
indicated in percent. Optical components (e.g., detector, source, and beam
splitter) as well as the card holder have an effect on the limits of the frequency
range.
●Wavenumber Accuracy Test: The wavenumber accuracy test verifies that the
frequency calibration of the spectrometer is within defined limits. Water vapor
is used for this test because it has extremely narrow bands, and the position of
these bands can be measured with a very high degree of accuracy.
Figure 40 shows a successful OQ test report.
OVP - OQ Test Protocol
Company :
Operator:
Instrument Ty pe:
Optics Configuration:
Accessory:
Instrument Serial Number:
Instrument Fir mwar e Ver sion:
OPUS/DB Version:
Overall Tes t Result
Test expires:
Test Date/Time:
Test Spectra Path:
Comment:
MILLIPORE
Default
Alpha Sample Compartment RT-DLaTGS ZnSe
Fibre 1 w ith: MIR, KBr, RT- DLATGS [Internal Pos.1]
Millipore Dir ect-Det ec t
M3
1.320_MDD_01 Oct 27 2011
OPUS 7.0 Build: 7, 0, 122 / DB: 7.0.105.1106
PA SSED
12/16/2012, 9:32:23 AM (GMT- 5)
12/16/2011, 9:32:23 AM (GMT- 5)
C:\Dire ct Detect\V alidation\Data\20111216\093223
Resolu tion Te st
Water Vapor Band:
Maxim um Reso lution:
1554.35 cm -1
2.00 cm -1 Measure d Resolution: 1.63 cm-1
Sens itivity Te st
Me asu re m ent Region , Start:
Minim um S/N:
2200.00 cm -1
2000
Meas urem ent Re gion, End:
Me asu re d S/N:
2100.00 cm -1
3446.07
Ener gy Dis tr ibution Tes t
Minim um Energy Value :
Minim um Energy Value :
Energ y at 7500.00 cm -1
5.00%
0.20%
0.01%
Energ y at 5000.00 cm -1
Ener gy at 500.00 cm- 1
9.14%
4.67%
Wavenu m be r Accuracy T es t Water Vap or
Expecte d Band:
Maxim um Deviation:
1576.130 cm -1
0.050 cm -1
Meas ured Band:
Me asu re d Deviation :
1576.129 cm -1
0.001 cm -1
Figure 40. OQ test report
13.4 Description of Instrument Validation Tests, continued
Direct Detect® Spectrometer User Guide
39
13.5 Running the Instrument Validation OQ and PQ Tests
1. Make sure card holder is empty.
2. To run OQ and/or PQ tests, click on the instrument validation icon .
3. When the instrument validation dialog appears, click on the Performance Qualification
(PQ) Run Test button (1 in Figure41). The test takes about 5 minutes.
Figure 41. Running OQ/PQ tests
2
1
Instrument validation
icon and OQ/PQ
Initiator
4. To run the Operational Qualification (OQ) test click on the Operational Qualification
Run Test button (2). The test takes about a minute to run.
For information on interpreting test failures, refer to section 12.4.
13.6 Storage
Store the Direct Detect® spectrometer and Assay-free cards in a clean, dry environment
at 15–30 °C.
Direct Detect® Spectrometer User Guide
40
14 Installation, Operation, and Process Qualification
Protocol (Optional)
NOTE: A 2 mg/mL protein standard (e.g., Thermo Scientific Pierce® or Bio-Rad® Bovine Serum
Albumin Standard ) is required for the protein verification test (section 14.5).
14.1 Installation Qualification Protocol (IQ)
NOTE: The installation qualification should be performed prior to making any local
modifications to the PC software.
1. Set up the system as instructed in section 6.5.1.
2. Verify that the Direct Detect® system starts the initialization sequence and that
successful completion is indicated by the status indicator light (5E in Figure 5) turning
green. Document results on the IQ/OQ/PQ Report Summary page found in section14.6.
14.2 Instrument Hardware Diagnostic Protocol
1. Access the Direct Detect® software by double-clicking on
the icon on the Netbook desktop.
2. Log into the instrument using the “Administrator” user name and password
“directdetect”. Once in the User Management screen, create a user name with
administrator privileges to run the OQ/PQ tests below. Refer to section 1.2.4, User
Management, in the Direct Detect® Software User Guide.
3. Click on the hardware diagnostics icon to open the hardware diagnostics dialog
(Figure42).
Direct Detect® Spectrometer User Guide
41
Figure 42. Instrument status
Hardware diagnostics icon
4. A green check mark indicates that each of the systems is functioning correctly. If
any systems do not have a green check mark, refer to section 5, Diagnostics, in the
Direct Detect® Software User Guide. Correct the problem, then click on the hardware
diagnostics icon to verify that all of the check marks are green.
Select the save button to save the hardware diagnostics report file to the
hard drive (C:\DirectDetect\Validation\Reports).
Select the print button to print the report to an installed printer.
5. Document results on the IQ/OQ/PQ Report Summary.
14.2 Instrument Hardware Diagnostic Protocol, continued
Direct Detect® Spectrometer User Guide
42
14.3 Operation Qualification Protocol (OQ)
1. Click on the instrument validation icon to open the instrument validation dialog box.
Figure 43. Instrument validation dialog
2
1
2. Click on the Operational Qualification (OQ) Run Test button (2). This starts a system
check that takes several minutes to complete. Run this test regardless of the test
expiration date. A report displaying the OQ test results will be generated and stored in
C:\DirectDetect\Validation\Reports. The report can be viewed and printed by clicking on
the button. Document results on the IQ/OQ/PQ Report Summary.
14.4 Performance Qualification Protocol (PQ)
1. After the OQ test has been successfully completed, run the PQ test by clicking on the
Performance Qualification (PQ) Run Test button (1). Run this test regardless of the
test expiration date. A report displaying the PQ results will be generated and stored in
C:\DirectDetect\Validation\Reports. The report can be viewed and printed by clicking on
the button. Document results on the IQ/OQ/PQ Report Summary.
14.5 Protein Measurement Verification
1. After successful completion of the OQ and PQ tests, the system is ready to verify that a
protein spotted on the card is equivalent to a standard.
2. Open a new vial of BSA standard.
3. Place the Assay-free card(s) on the spotting tray or other clean surface. Select an
appropriate buffer blank, preferably the same solution in which the BSA standard has
been prepared. The majority of commercially available BSA standards use NaCl (with
NaN3) as a buffer; check manufacturer specifications. Pipette exactly 2 µL of the buffer
solution to the default blank position on the card (position1).
Direct Detect® Spectrometer User Guide
43
14.5 Protein Measurement Verification, continued
4. Pipette exactly 2 µL of sample to sample positions 2 through 4. Card positions can be
labeled if desired.
5. In the Direct Detect® Software main screen, complete the measurement setup as
indicated in Figure 44. Choose NIST BSA AM1 from the protein drop down menu and
check the Dry sample card box. Sample location 1 should be blue, indicating that the
buffer solution is in this location. Sample locations 2–4 should be green, indicating
that there is a sample to be measured.
Figure 44. Sample information
6. Follow the instructions in section 10.2 for inserting the sample card into the
instrument, then click on the button.
7. The sample measurement results are displayed on the Last Sample Result tab of the
main screen.
8. Measure the BSA standard concentration using a UV reader at 280 nm.
DO NOT use standard curves built into the UV reader. Use the sequence-based
extinction coefficient 42,925 M-1cm-1 or ABS 1 mg/mL = 0.646. Zero the UV instrument
with the correct buffer and read absorbance at 280 nm.
9. Manually calculate the correct concentration using the sequence-based numbers. The
system passes the protein verification test if the results from the UV reader and the
Direct Detect® system are within 10% of one another. Document results on the
IQ/OQ/PQ Report Summary.
NOTE: The Direct Detect® system comes pre-loaded with standard curves that
were generated from phosphate-buffered saline (PBS). Analysis from buffers
like NaCl with NaN3 delivers accurate quantification if the measurement is
performed in a room with humidity level below 35%. Humidity levels above
35% will affect sample drying, resulting in artificially increased concentration
estimates.
Direct Detect® Spectrometer User Guide
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14.6 Direct Detect® System IQ/OQ/PQ Report Summary
Reference
Section
Checklist Result
Pass Fail
14.1 Verify power and cable connections
● Power adapters are connected to Netbook computer and
Direct Detect® instrument
● Verify connection between Netbook computer and Direct
Detect® instrument (cross over cables)
Verify Software communication
● Direct Detect® Software launches successfully
● Login successful
● Instrument detected
14.2 Instrument Hardware Diagnostic
● All functions have green check mark
14.3 System OQ
● OQ test report indicates Overall Test Result = PASSED
14.4 System PQ
● PQ test report indicates Overall Test Result = PASSED
14.5 Verification of Protein Measurement
● Direct Detect® measurement and UV reader measurement
are within 10% of one another for a 2 mg/mL BSA standard.
Testing completed by:
Date:
Direct Detect® Spectrometer User Guide
45
15 Specifications
General
Dimensions 22 cm wide x 30 cm deep x 15.2 cm high (including tray)
(8.7 x 11.8 x 6.0 in.)
NOTE: Overall height with Netbook computer opened will vary,
but is typically less than 40.6 cm (16 in.).
Weight (approximate) 7 kg (15.4 lb)
Power consumption Maximal: 100–240 VAC, 50–60 Hz (max 2 A)
Electronics Class 1 laser product (contains 850 nm laser diode)
Environmental conditions Operational temperature range: 18–35 °C (64–95 °F)
Temperature variation: max. 1 °C/h and max. 2 °C/day
Humidity (non-condensing): less than 35% (relative humidity)
Humidity variation: less than 3%/hr
Installation site: in a closed room, max. 2,000 m above sea level
Installation overvoltage category: II
Pollution degree: 2
Safety regulations Complies with EN/IEC 61010-1/08.2002 (Safety requirements for
electrical equipment for measure ment, control and laboratory use)
EMI regulations Complies with EN/IEC 61326/05.2004 (Electrical equipment for
measurement, control and laboratory use - EMC requirements)
Performance
Spectral range 6,000–500 cm
-1
Resolution Better than 2 cm
-1
Wave number accuracy Better than 0.05 cm
-1
Wave number reproducibility Better than 0.01 cm
-1
Photometric accuracy 0.1% T
Optics
Design Sealed optics housing, convective desiccating
IR Source Low voltage, air cooled, can be replaced
(lifetime 44,000 operating hours)
Computer Specifications
Operating System Microsoft® Windows® 7
Software Framework Microsoft®.NET Framework 2.0
Screen Resolution 1,024 x 600 or greater
Ethernet Cable Crossover, 10Base-T (10 or 10/100 MBit) with RJ45 connectors
Processor Intel® Pentium® III, 800 MHz or greater
Random Access Memory (RAM) 2 GB
Graphics Card XVGA, 256 Mb RAM, 32-bit color depth, Open GL, GDI+
Direct Detect® Spectrometer User Guide
46
16 Ordering Information
This section lists catalogue numbers for the Direct Detect® spectrometer and replacement parts. See
Technical Assistance section for contact information. You can purchase these products on-line at
www.millipore.com/products.
Product Description Cat. No. Qty/Pk
Direct Detect® Spectrometer
Direct Detect® Spectrometer (includes instrument with power
adapter*, Dell® Netbook computer with power adapter*,
Ethernet crossover cable, Netbook computer stand, 50-pack
of Assay-free cards, spotting tray, spare desiccant bag, Torx®
screw driver, Quick-Start Guide)
* Power adapters are configured for multiple countries
DDHW00010-WW
1
Direct Detect® Spectrometer
Replacement Parts/Accessories
Direct Detect® Assay-free Cards Multiple colors
Multiple colors
Green
DDAC00010-4P
DDAC00010-8P
DDAC00010-GR
50 x 4
50 x 8
50
Spotting Tray DDMISC0010-ST 5
Netbook Stand DDMISC0010-NS 1
Direct Detect® IR Source** DDSP00010-LA 1
Direct Detect® Desiccant Bag** DDSP00010-DE 1
** Contact Technical Service to order
(see Technical Assistance section)
Direct Detect® Spectrometer User Guide
47
17 Technical Assistance
For more information, contact the office nearest you. In the U.S., call 1-800-MILLIPORE
(1-800-645-5476). Outside the U.S., go to our web site at www.millipore.com/offices for up-to-date
worldwide contact information. You can also visit the tech service page on our web site at
www.millipore.com/techservice.
18 EC Representative
Millipore (UK) Ltd.
Fleming Road, Kirkton Campus
Livingston EH54 7BN
UK
19 Standard Warranty
The applicable warranty for the products listed in this publication may be found at
www.millipore.com/terms (within the “Terms and Conditions of Sale” applicable to your
purchase transaction).
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