SERVA HPE™ ScreenPicker INSTRUCTION MANUAL Semi-manual spot picking system

SERVA HPE™ ScreenPicker INSTRUCTION MANUAL Semi-manual spot picking system
INSTRUCTION MANUAL
SERVA HPE™ ScreenPicker
Semi-manual spot picking system
(Cat. No. HPE-SP1)
SERVA Electrophoresis GmbH  Carl-Benz-Str. 7  D-69115 Heidelberg
Phone +49-6221-138400, Fax +49-6221-1384010
e-mail: [email protected]  http://www.serva.de
Contents
1. Introduction
2
2. Features of the Screen Picker
3
2.1. Specifications
3
2.2 Description of the ScreenPicker
4
2.3. Packing List
5
3. Installation
6
4. Operation
6
4.1. Image Viewer software manual
4.1.1. Automatic mode
4.1.2. Manual mode
4.1.3. Mixed mode (automated with additional manual spots)
7
13
15
5. Software re-installation
18
5.1. General comment on software protection
18
5.2. Installation from Boot menu
19
5.3. Installation from USB backup key
19
5.4. License key installation
20
6. Care and maintenance
20
7. Picker.ini configuration**
20
8. Closing and Shutdown procedure
21
9. Important comments
21
10. Legal Information
21
11. Order Information
22
Vers. 11/12
1
8
1. Introduction
Manually picking from a fluorescent 2D electrophoresis gel is a challenge without an
automated picking robot. It requires to post stain the gel in order to visualize the
spots. Many classical staining methods interfere with mass spectrometry or if not,
have lower sensitivity than fluorescent staining.
The SERVA HPE™ ScreenPicker is a device to semi-manually pick fluorescent 2D
gels (DIGE, LavaPurple, Sypro Ruby, Flamingo…) without the need of post staining
and/or sophisticated robot and software. It consists of an embedded screen
connected to a small computer to display the scanned image of the gel, an XY
carriage to guide the picker accurately over the spot position and a software which
can read images and picking lists from most evaluation packages. The ScreenPicker
is adapted for gels on 3 mm glass plates (fluorescent markers placed on 3 corners
before scanning) and HPE™ gels on non-fluorescent plastic backing (punched holes
performed on 3 corners before scanning). The scanned image is projected under the
gel. The provided software allows performing a fine-tuning between the size of the
displayed image with a mouse and the size of the real gel with the help of markers or
punched holes used as landmarks. An XY carriage is used to guide the manual
picker over the target to be excised and to prevent parallax errors. The ScreenPicker
guides visually the user to place the picked plug in the right well of the microtiter plate
using the file generated by the evaluation software. At the end of the process a file
containing spot numbers and microtiter plate plug positions is generated for use by
digester robot.
Because it clearly marks the spots to pick on the image, the ScreenPicker can also
be used for Coomassie® blue or silver stained gels, increasing the picking accuracy
and making the process entirely error free.
Another interesting application is the picking of protein spots from 2D gels after the
immunological visualization of specific spots by western blotting: The scanned image
of the blot membrane is projected under the gel, which contains still a great portion of
the proteins after the electro-transfer.
The files from the evaluation software are transferred to the Netbook PC via an USB
stick. The special Crunchbang (Debian) Linux installation on the Netbook PC is
minimalistic. The Netbook PC setup is not designed for any other purpose than
picking spots.
2
Benefits
• Space saving: easy to store on lab bench; dimensions length, width, height
compared to huge automatic picker.
•
Time saving, accuracy, easy to use, no need for specific training, no need for
manual corrections of the gaps always encountered with automatic pickers.
Full workflow integration with generation of picking-digester list. Ready within
seconds.
•
Costs saving: no need for expensive instrument or maintenance contract.
•
Error free picking with help of pick list and plate well identifier.
•
Flexibility: initialization file defining visual appearance and requirements of
digestion robot like input file and reserved positions on plate. Reads any
image file format or picking list.
•
Safety: no transilluminator or dangerous UV sources.
•
Security: Linux avoids transfer of viruses by USB memory keys. PC and
Software protected.
2. Features of the Screen Picker
2.1. Specifications
The system is mainly designed for film-backed HPE™ gels and gels on 3 mm glass
plates.
Screen box
Dimensions
Weight
Features
55 x 36 x 8 cm
7 kg
Visible light for safe usage
Leakage-free
Flat screen protected by glass platen
Maintenance-free
Precise XY carriage
Robust default picker head 1.5 mm diameter
Voltage
Power Consumption
AC 90V to 264V; 60 Hz
Power rating 1.2 A at start (protected by fuse in switch )
37 W nominal
10° C to 40° C
Relative Humidity: 20% to 85%
Operating Temperature:
Environment
3
Computer hardware
Netbook PC 10.1“, 1024 RAM, 250 GB HD,
AC 90V to 260V; 50-60 Hz. 30 W maximum.
Linux-based software environment, virus-safe
Connection to screen box via monitor cable
Cable free mouse
USB stick for data transfer
ImageViewer software
Flexible: reads any image or pick list format
Variable configuration for plug disposal in X or Y direction;
reserved positions in MTP; spot cross hair size on screen.
Manual spot selection saved to file.
Function icons for easy handling
Adjustable image contrast
“SYSTEM BACKUP” USB stick
Kolourpaint tool
Paint program for marking and documenting manually
selected spots when working without image analysis
software.
2.2 Description of the ScreenPicker
Fig. 1: The complete Screen Picker system
4
Fig. 2: Screen box setup
2.3. Packing List
•
•
•
•
•
•
Screen picker box
Y- rail and carriage for picking head
One Touch 2D gel spotpicker 1.5 mm (Cat. No. P2-D15)
Power cord
VGA Monitor cable
Netbook PC with power supply and power cord
• Cordless mouse
• USB key for data transfer
• USB key for system backup
•
•
Dust cover (Cat. No. HPE-SP02)
450 Fluorescent stickers (5 mm diameter, Cat. No. HPE-FS05)
• Waterproof tape
• Instruction manual
5
3. Installation
1. Unpack the ScreenPicker and place it on a flat surface with the label facing
you.
2. Unpack the Y rail and insert it from the left or right of the two parallel X rails,
with the double guiding insert facing you..
3. Insert the carriage with picker support on the Y rail. You can decide to have
the picker head on the left or on the right of the Y rail.
4. Connect the provided VGA (15 pins sub-D) cable on the computer and the
mains cable to the unit. Connect computer to mains via power supply
according to computer installation guide.
5. Start the computer, the software running under Linux will load immediately
after security check.
6. Use USB keys or drive to transfer the image and picklist of the gel to pick.
Format allowed are Tiff, .gel, mel, img, jpg, png, bmp, gif, 8, 12, 16 bits or
more... Conversion is fully automated.
7. Pick list can be imported from major evaluation software packages; format
must be ASCII comma delimited.
8. The system is delivered fully configured but can be customized at installation
by editing the picker.ini file located in ../param folder (see section 4.1.1.).
4. Operation
The ImageViewer software delivered on the Netbook computer is running under a
special version of Linux (CrunchBang Linux). .
Fully charge the computer before first use following computer installation guide. On
start up, a user name and a password must be provided to access the safe
environment. Unless it has been changed during installation, the username is
“picker” and password is “spot”; both are case sensitive.
In case of software failure, follow procedure in chapter 5, “Software re-installation”.
Important before Scanning the Gels:
•
Remember to put fluorescent markers (adhesive white dots on the glass plate
on 3 edges before casting the gel in case of traditional vertical gels. (Fig. 3)
•
While running HPE gels, either use small scissors to make V cuts on 3 edges
(Fig. 3) or the fluorescent adhesive dots.
6
Fig. 3: Marker positions on plastic-backed gels and glass plate gels.
Switch on the device using the button on the left side (Fig. 2). Then switch on the
Netbook computer
The booting process takes approximately 30 sec.
4.1. Image Viewer software manual
ImageViewer software is automatically loaded after boot. If the program has been
closed, a right click with the mouse opens a dialog box, click on first menu item
<ImageViewer>.
An array of icons (figure 4) makes handling easy:
Fig. 4: Imageviewer icons on the screen.
Please notice that the direction of filling double arrow is just an indicator, you can change the
direction via picker.ini file in chapter 7.
7
4.1.1. Automatic mode
In automatic mode, the loaded picking list is entirely driving the process by
highlighting one spot at a time by a cross hair indication and the corresponding well
position to release the picked plug.
1. Choose the file name of the gel to pick from the browser window popping up
as per Fig. 5, and click <Open> It is also possible to choose an image file
using camera icon on menu bar (The USB key used to transfer files is labelled
. “xGb filesystem “ on the folder list ).
2. A second dialog box opens to choose the corresponding picking list . Select
the file with the mouse and press <Open> (Fig. 6) or press <Cancel> to pick
without pick list (see Manual mode 3.1.2) It is also possible to choose picking
file using printer icon on menu bar.
The folder containing image and picking list files will be saved automatically at the
end for the next run.
Choose “Ettan™ picker” as picking robot in Imaging software to generate the picking
list
Fig. 5: Choosing image from gel
to pick
Fig. 6: Choosing corresponding picking list
As the last position used in the microtiter plate is saved at the end of the run , before
starting, you have to choose between filling the next empty position on an existing
plate or restart at the default position on a new plate (the default position is defined in
the initialization file). This feature allows the user to pick several gels using the same
microtiter plate. (Fig. 7.)
Fig. 7: Continue to fill existing plate or choose a new one
8
3. Lay your gel on the picker window, gel side up. Start by positioning one V cut
or marker on the corresponding image position. In general, it is not necessary
to fix the gel but if needed, use the tape provided.
4. Using mouse, enlarge the image by dragging right and bottom sides to get the
2 other markers properly coincide with the real gel (holes or markers); Use the
XY carriage without the picker to avoid any parallax distortion. Lock the image
position and size by pressing locker icon; icon becomes red.
5. The 96-wells microtiter plate used to collect the polyacrylamide plugs is placed
on the bottom right corner of the screen picker. There is only one way to
position the plate due to the special design (Fig. 8).
Fig. 8: Positioning the microtiter plate
17. Image contrast is adjusted using icons on menu bar (Fig. 4) in section 4.1.
This icon opens dialog box enabling the
choice of contrast steps:
This icon increases contrast by number
of steps defined above
This icon decreases contrast by number
of steps defined above
6. ImageViewer is running in Auto mode (icon A of Fig. 4), the software will
display the first spot to pick with a crosshair on the image and it will also
highlight the well position to use by means of the plate coordinates and a red
spot visible underneath the plate at the correct well position. (Fig. 9,10).
9
7. Move the carriage to the pick position like on Fig. 11; move Y rail by moving
left-right only the X carriage located on front side (Fig. 12, 13). Immobilize the
XY carriages by rotating clockwise 1/4 turn the black knurled thumb screw
knobs, introduce the picker into the 2 holes of the picker head (Fig. 14).
Fig. 9: Plug position highlighted below
the Microtiter plate and indicated by
arrows (for black MTPs).
Fig. 10: Positions in black are reserved
for calibration, the display shows the plate
Nr 3 position D4 to put spot 1830
Fig. 11: Crosshair generated on spot to
pick
Fig. 12: Moving X carriage from front side
10
Fig. 13: Align the carriage over the spot
using XY lines and the spot crosshair.
Fig. 14: Use black knurled thumb
screw knobs to lock carriages.
8. Pick the spot by pushing downwards the black tube of the picker to close the
jaws. Rotate the picker one full turn to detach the plug from glass or plastic
(Fig. 15).
9. Remove the picker from the carriage and dispense the plug into the microtiter
plate at the indicated / highlighted position (Fig. 16). Carefully rinse your picker
after each spot to avoid cross-contamination.
Fig. 16: Placing the spot plug into plate
ready for digestion.
Fig. 15: Picking the spot.
10. Use arrows keys on the menu bar to continue with next spot.
11
11. When you reach the last position on the plate a message invites you to put a
new plate :
The microtiter plate number will be incremented on
screen and in file.
12. At the end of the process, a window pops up
saying “You are on the last spot”; click “OK” and
...
13. At that stage you can switch to
manual mode to define new spots
to pick or not; If you select “Yes”,
refer to 3.1.2 step 7 If you answer
“No”, continue with step 15.
14. Click the green arrow icon on the menu bar to save the picking list as a file for
digester; The created file will contain spot numbers, spot positions, microtiter
plate number (default=1) and the plate well position for each spot picked (e.g.:
2C11= plate 2 raw C column 11). The software automatically identifies the
positioning markers (IR1 & IR2) used by Ettan™ picker (GE Healthcare) and
remove these from picking process and saved file.
The file name and folder are entered via the dialog box. By default, the file will be
saved in the folder used to load image and pick list but you can save it in any other
folder by selecting <Browse for other folders> (Fig. 17). An example of the saved list
is visible on Fig. 18.
12
Fig. 17: Browsing between folders.
15. At that stage, you can switch to the following gel to
pick by pressing the Camera button on menu bar.
You are now jumping again at step 1 of this
chapter. If you didn’t save the file for the digester
yet, you will get a reminder ...
If you do not save the pick list, the last well
position used is not saved!
16. Before removing the USB key, press the
USB icon on the title bar to close files and
eject key or drive; remove your key only
when you see this message:
4.1.2. Manual mode
The manual mode gives the scientist the freedom to decide which spot he wants to
pick. The spot on the gel will be identified later by his XY coordinates in the image
and will be reported as such in the digester file. There is no limit to the number of
manual spots to pick. ImageViewer allows working in mixed mode: automatic mode,
switching to manual and back to automatic (See section 4.1.3.).
We know by experience that researchers are sometimes interested to pick additional
spots for instance in order to identify a train of different glycosylations on a gel near a
spot identified by the imaging software as a statistically relevant difference. In case of
large spots, it could also be a good idea to pick more than once; this may help to
identify multiple proteins present in that spot. Most imaging software give the
possibility to only pick one time per spot. With automated picker, it is also not so easy
to add more positions to the picking list or to change the picker head for a larger one.
Screen Picker gives you all these features. If you have a faint spot, why not choosing
a picker of 3 mm instead of 1.5 mm diameter or picking more than once in the spot!
13
1. Choose the file name of the gel to pick from the browser window popping up as
per Fig. 5, and click <Open> It is also possible to choose an image file using
camera icon on menu bar. (The USB key used to transfer files is labelled . “xGb
filesystem “ on the folder list)
2. A second dialog box opens to choose the corresponding picking list . Press
<Cancel> to pick without pick list.
The folder containing image and picking
list output files will be saved automatically
at the end for the next run.
As the last position used in the microtiter plate is saved at the end of the run ,
before starting, you have to choose between filling the next empty position on
an existing plate or restart at the default position on a new plate (the default
position is defined in the initialization file). This feature allows the user to pick
several gels using the same microtiter plate (Fig. 7).
3. Lay your gel on the picker window, gel side up. Start by positioning one V cut or
marker on the corresponding image position. In general, it is not necessary to fix
the gel but if needed, use the tape provided.
4. Using the mouse, enlarge the image by dragging right and bottom sides to get
the 2 other markers properly coincide with the real gel (holes or markers); Use
the XY carriage without the picker to avoid any parallax distortion. Lock the
image position and size by pressing locker icon; icon becomes red.
5. The 96 wells microtiter plate used to collect the polyacrylamide plugs is placed on
the bottom right corner of the screen picker. There is only one way to position the
plate due to the special design (Fig. 8).
6. Image contrast is adjusted using icons on menu bar (Fig. 4 in section 4.1)
7. Click on the image at the position you plan to pick a spot, a dialog box will pop
up: click “No”, to reselect another position; click “Yes” will validate the spot to
pick.
8. The spot number will be incremented to Mxx (starting with M1)
and displayed on the image, Move the carriage to the pick
position like on Fig. 13; move Y rail by moving left-right only the
X carriage located on front side (Fig. 12). Immobilize the XY
carriages, introduce the picker into the 2 holes of the picker
head.
9. Pick the spot by pushing downwards the black tube of the
picker to close the jaws. Rotate the picker one full turn to
14
detach the plug from glass or plastic (Fig. 15).
10. Remove the picker from the carriage and dispense the plug into the microtiter
plate at the highlighted position (Fig. 16). Carefully rinse your picker after each
spot to avoid cross-contamination.
11. Continue with step 7 for the next manual spot
12. Terminate the picking process by clicking the green arrow icon on the menu bar
to save the picking list as a file for digester; The created file will contain spot
numbers (M1 to Mx), spot X,Y positions, microtiter plate number (default=1) and
the plate well position for each spot picked (e.g. 2C11= plate 2 raw C column 11).
The file name and folder are entered via the dialog box. By default, the file will be
saved in the folder used to load image but you can save it in any other folder by
selecting <Browse for other folders> (Fig. 17).
13. At that stage, you can switch to the following gel to pick by
pressing the Camera button on menu bar. You are now
jumping again at step 1 of this chapter. If you did not save
the file for the digester yet, you will get a reminder ... If you
do not save the pick list, the last well position used is not
saved!
14. Before removing the USB key, press the USB icon on the title bar to close files
and eject key or drive; remove your key only when you see this message :
4.1.3. Mixed mode (automated with additional manual spots)
In mixed mode, the loaded picking list is entirely driving the process by highlighting
one spot at a time and the corresponding well position to release the picked plug. At
any time, the user can decide to switch to manual for choosing additional spots to
pick; the automated process will continue and the manual spots chosen will be added
at the end of the process. The advantage of this mode is for the user to make
multiple picks into one large spot or to pick adjacent spots which seems to be related
to spots chosen by the evaluation software.
1. Choose the file name of the gel to pick from the browser window popping up as
per Fig. 5, and click <Open> It is also possible to choose an image file using
camera icon on menu bar. (The USB key used to transfer files is labelled . “xGb
filesystem “ on the folder list).
15
2. A second dialog box opens to choose the corresponding picking list. Select the
file with the mouse and press <Open> (Fig. 6) or press <Cancel> to pick without
pick list (see Manual mode 4.1.2) It is also possible to choose picking file using
printer icon on menu bar. - The folder containing image and picking list files will
be saved automatically at the end for the next run. - Choose “Ettan™ picker” as
picking robot in Imaging software to generate the picking list - As the last position
used in the microtiter plate is saved at the end of the run, before starting, you
have to choose between filling the next empty position on an existing plate or
restart at the default position on a new plate (the default position is defined in the
initialization file). This feature allows the user to pick several gels using the same
microtiter plate. (Fig. 7).
3. Lay your gel on the picker window, gel side up. Start by positioning one V cut or
marker on the corresponding image position. In general, it is not necessary to fix
the gel but if needed, use the tape provided.
4. Using mouse, enlarge the image by dragging right and bottom sides to get the 2
other markers properly coincide with the real gel (holes or markers); Use the XY
carriage without the picker to avoid any parallax distortion. Lock the image
position and size by pressing locker icon; icon becomes red.
5. The 96 wells microtiter plate used to collect the polyacrylamide plugs is placed on
the bottom right corner of the screen picker. There is only one way to position the
plate due to the special design (Fig. 6).
6. Image contrast is adjusted using icons on menu bar (Fig. 4 in section 4.1 )
7. ImageViewer is running in Auto mode (icon A of Fig. 12), the software will display
the first spot to pick with a crosshair on the image and it will also highlight the
well position to use by means of the plate coordinates and a red spot visible
underneath the plate at the correct well position (Fig. 9,10).
8. Move the carriage to the pick position like on Fig. 13; move Y rail by moving leftright only the X carriage located on front side (Fig. 12). Immobilize the XY
carriages , introduce the picker into the 2 holes of the picker head.
9. Pick the spot by pushing downwards the black tube of the picker to close the
jaws. Rotate the picker one full turn to detach the plug from glass or plastic (Fig.
15).
10. Remove the picker from the carriage and dispense the plug into the microtiter
plate at the highlighted position (Fig. 16). Carefully rinse your picker after each
spot to avoid cross-contamination.
11. At any time, switch to manual mode by depressing icon A on menu bar, the icon
will turn white in manual mode and the message bar at the bottom of the image
window will display:
16
12. Click on the image at the position you plan to pick a spot, a
dialog box will pop up: click No, to reselect another position;
click Yes will validate the spot to pick. Perform this step as
many times you want to pick new manual spots. The
software will remember the manual coordinates of new spots
and will guide you at the end of the automatic process.
13. Use arrows keys on the menu bar to continue with next spot. ImageViewer is now
jumping to next automatic spot to pick; at the end of the spots from the picking
list, the plate display will change to show the manual spots starting with M1 until
Mxx. The process can be interrupted as many times as you want by switching
back again to step 9.
14. When you reach the last position on
the plate a message invites you to put
a new plate: The microtiter plate
number will be incremented on screen
and in file.
15. At the end of the process, a window pops up
saying “You are on the last spot”; click OK and ...
16. At that stage you can still add manual spots (see
step 10). After each manual spot added, you will get the message as in step 13.
17. Terminate the picking process by clicking the green arrow icon on the menu bar
to save the picking list as a file for digester; The created file will contain spot
numbers (automated and manual at the end), spot positions, microtiter plate
number (default=1) and the plate well position for each spot picked (e.g. 2C11=
plate 2 raw C column 11). The software automatically identify the positioning
markers (IR1 & IR2) used by Ettan™ picker and remove these from the picking
process and from the saved file. The file name and folder are entered via the
dialog box. By default, the file will be saved in same folder than the folder used to
load image and picklist, but you can save it in any other folder by selecting
<Browse for other folders> (Fig. 13).
An example of the list saved is visible in Fig .18.
17
Fig. 18: Example of pick list saved for the digester.
18. At that stage, you can switch to the following gel to pick
by pressing the Camera button on menu bar. You are
jumping again at step 1 of this chapter. If you didn’t save the
file for the digester yet, you will get a reminder ...
If you do not save the pick list, the last well position used is
not saved!
19. Before removing the USB key, press the USB
icon on the title bar to close file and eject key or
drive; re-move your key only when you see this
message:
5. Software re-installation
5.1. General comment on software protection
ImageViewer application is protected by a dual level security encrypting key. You
cannot install the package on another machine than the computer which is delivered
with the system. We sell additional pre-installed computers in case of need. Please
ask your distributor for a quotation.
There are two ways of re-installing the software on the computer.
18
5.2. Installation from Boot menu
•
Always attach the computer to mains supply before continuing.
•
At boot, the GRUB Linux loader is visible. If you do not do anything, the computer
will automatically boot and load the software after 20 sec. Pressing the “Enter”
key forces the system to boot immediately.
•
In case of boot failure identified by a message on screen, switch off the computer
by pressing the Power on switch for a few seconds until computer power goes
off.
•
Power computer on again and when you see the GRUB Linux loader, move down
using arrow keys to the last line labelled : “Software installation (use only in case
of boot failure)”
•
A dialog will ask you to confirm your choice, answering “No” will start the process
again, “Yes”, will initiate the complete re-installation at factory defaults.
•
Follow instructions on screen until end of the process..
DO NOT POWER OFF COMPUTER DURING INSTALL PROCESS.
5.3. Installation from USB backup key
A special USB key containing a full backup of operating system and application is
included with the package in case the software must be reinstalled due to computer
crash.
This key is labelled “SYSTEM BACKUP” In case you cannot find back the USB key
included in the original package, please ask your distributor for a copy provided at
production cost price.
•
Always attach the computer to mains supply before continuing.
•
If the process detailed in 5.2 has failed, please switch off your computer by
pressing power on switch for a few seconds until computer power goes off.
•
Attach your “System Backup” USB key to any USB port and switch computer
on again.
Follow all instructions on screen until you get the final message inviting you to
remove the USB key and reboot.
The system must be up and running again at factory defaults.
DO NOT POWER OFF COMPUTER DURING INSTALL PROCESS
19
5.4. License key installation
In case the license key must be re-installed, use contact details (phone, email)
provided on screen to fix the issue on your system.
6. Care and maintenance
Immediately after use, wipe the glass plate with a clean tissue and
water (can use 0.1% SDS). Never use any other solvent than methanol
or 50:50 ethanol.
Always make sure that the rails are clean; eventually clean them with
distilled water; never use grease, oil or any lubricant. For smooth
travelling of Y rail, use only X carriage located on front side (Fig. 8 bis).
Rinse properly OneTouch spot picker between each plug to avoid crosscontamination.
7. Picker.ini configuration**
There is a configuration file located in /home/picker/imageViewer/param. We do not
recommend to change the X and Y offset for the 96 wells plate, size and gap of the
spot crosshair and plateW, plateH (this has been optimized for each machine). You
can change the default start, the reserved positions on the microtiter plate. Some
users like to keep wells for standards or calibration. It is also possible to adjust the
contrast changing speed, the plate geometry and the way of filling the plate.
Procedure:
- Double click on file picker.ini file to
open it.
- Enter the position to start in the line
“start= “
- Enter positions to skip in line “reserve”
without using space between values.
- Contrast is the default contrast change
speed (0.5 steps)
- Way is the plate filling direction; V
means A1,A2,A3..., H means
A1,B1,C1,D1... (An orange double arrow
indicating the direction, is visible on image window
title bar (Fig. 12)).
- PlateR is the plate geometry; 0 is
normal, 1 is inverted
- Save the file after edition by clicking “Save” in menu bar.**
**: ask help from your IT department or Proteomics Consult if questions
20
8. Closing and Shutdown procedure
•
Close ImageViewer by pressing the <Close> icon on image title bar (Fig. 4).
•
Press Computer power button once to initiate shutdown procedure or right
mouse click and choose “Shutdown”
•
Shutdown screen picker using Power button on the left (Fig. 2).
9. Important comments
The special Crunchbang (Debian) Linux installation is minimalistic. The system is not
designed for any other purpose than picking spots. In case of network installation,
please communicate network details before installation and inform IT people in
advance to help configuring the system on the network at the install. We do not
recommend using file transfer via network to avoid support problems. In case of
network link, we truly recommend to subscribe a maintenance contract with the
support team. Please ask your distributor for a quotation.
ImageViewer is fully protected from changes or copy via dual-level encrypting license
key. The delivered package on an USB key cannot be installed on another computer.
Please contact your distributor
10. Legal Information
Screen Picker is covered by Intellectual Property rights assigned to Proteomics
Consult for the design and the hardware, and to Alltux for the software
(http://www.alltux.be )
21
11. Order Information
Product
HPE™ ScreenPicker
Plexiglass protection shield, 50 cm wide
OneTouch 2D gel spot picker, 1.5 mm diameter
OneTouch 2D gel spot picker, 3.0 mm diameter
Screen Picker Protection Cap
Fluorescent Stickers (450 pcs., Ø 5 mm)
Cat. No.
HPE-SP1
HPE-PS50
P2-D15
P2-D30
HPE-SP02
HPE-FS05
Important
Specifications, software bundles, and accessories are subject to change without
notice. Not responsible for typographic errors.
Related products
Product
Trypsin NB Premium Grade, MS approved
Trypsin NB Sequencing Grade, modified
Endoproteinase Glu-C (V8), MS approved
22
4 x 25 µg
4 x 25 µg
4 x 25 µg
Cat. No.
37384.01
37283.01
20986.01
Appendix A
When spots are picked from a gel without the use of a picking list, there is a tool to
annotate the image in advance with the spots to pick. This feature is useful if you do
not have an imaging software application, or if the number of gels is anyway too
small to be analyzed by such a software.
1. Scan the gel with the markers as described in chapter 3.
2. Load your image on the USB key and insert the key into the picker
computer. The file manager will pop up to display the folders on the USB
key.
3. Double click on the image to modify. The application Kolourpaint is loaded
with the image.
4. You can zoom in and out or modify contrast using the buttons “Zoom in”
“Zoom out” and “Enhance Image” (Fig. A.1)
Fig. A.1: Kolourpaint Zoom and contrast buttons
Fig. A.2: Kolourpaint painting buttons
5. From the left toolbox, choose the ellipse object, the line width and from the
bottom, you can select the colour. Click and drag on the gel around the
spot you will select for later picking. (Fig. A.2)
6. It is possible to write a label annotation near the spot for easy identification.
The font type & size are selected via the buttons on top (Fig. A.1.) The font
colour is selected via the colours on the bottom. Select the text tool via the
left toolbox; you can now see a new icon to choose between transparent
background or solid. Choose transparent, click and drag near the spot and
write the annotation. (Fig A.3)
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7. Continue at step 5 again for additional spots.
8. When the selection is finished, save the file via the menu bar : File->Save
Image As ; Image format can be chosen via Filter, but according to original
format, the software will propose the closest possible. See Fig. A.4.
Fig. A.3: Kolourpaint writing tools
Fig. A.4: Kolourpaint Saving image at the end
9. The modified image can now be imported into the ImageViewer software
for picking. Please notice that the palette colour of the image is modified to
show the same colour like the one you used during image annotation. A
good advice will be to pick the spots in manual mode in the same order as
the spots annotations. The spots in the final saved digester file will
correspond to the image (Fig. A.5 and A.6).
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Fig. A.5: Kolourpaint image at saving
Fig. A.6: ImageViewer image during picking of spot 1
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