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GLYCO DOC GEL IMAGING SYSTEM - Bio-Rad

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<strong>GLYCO</strong> <strong>DOC</strong><br />

<strong>GEL</strong> <strong>IMAGING</strong> <strong>SYSTEM</strong><br />

INSTRUCTION MANUAL<br />

Catalog Numbers<br />

170-6555<br />

170-6556<br />

170-6557<br />

170-6558<br />

170-6559


Table of Contents<br />

TABLE OF CONTENTS<br />

Safety<br />

Section 1.0 Installation and Startup ...............................................................................................1<br />

1.1 Unpacking.......................................................................................................................2<br />

1.2 Set-up the Computer and Imager...................................................................................2<br />

1.2.1 Installing the Camera Circuit Board in a Computer..............................................2<br />

1.2.2 Setting Up the System..........................................................................................4<br />

1.2.3 Installing the Glyco Doc Software ........................................................................6<br />

1.3 Verify system Operation .................................................................................................7<br />

1.3.1 Starting the Glyco Doc Software ..........................................................................7<br />

1.3.2 Acquire an Image using the Test Plate.................................................................8<br />

1.4 Image Analysis Using the Image Provided with your System........................................10<br />

1.5 Preparing for Gel Imaging ..............................................................................................13<br />

1.6 Example of a Standard Lane Analysis ...........................................................................14<br />

Section 2.0 Software Reference......................................................................................................15<br />

2.1 Files Menu......................................................................................................................15<br />

2.2 Image Menu ...................................................................................................................18<br />

2.3 Bands Menu ...................................................................................................................21<br />

2.4 Options Menu .................................................................................................................24<br />

2.5 Window Menu.................................................................................................................25<br />

Section 3.0 Theory of Operations...................................................................................................28<br />

Section 4.0 Maintenance And Troubleshooting.............................................................................29<br />

4.1 Replacing the UV Light ..................................................................................................29<br />

4.2 Cleaning the Excitation Filter .........................................................................................31<br />

4.3 Preparing Gel plates.......................................................................................................32<br />

4.4 Creating a New bright field.............................................................................................32<br />

4.5 Imager Troubleshooting..................................................................................................34<br />

4.6 Software Troubleshooting...............................................................................................35<br />

4.6.1 Memory Problems.................................................................................................35<br />

4.6.2 Windows ...............................................................................................................35<br />

Appendix A. Initialization File ...........................................................................................................36<br />

A.1 Window Display Defaults................................................................................................36<br />

A.2 Standard Lane Option Defaults......................................................................................36<br />

A.3 Colors .............................................................................................................................37<br />

A.4 Band Descriptions ..........................................................................................................38<br />

A.5 Last Exposure Time........................................................................................................39<br />

A.6 Bright File Locations.......................................................................................................39<br />

A.7 Default Data Locations...................................................................................................39<br />

Appendix B. Imager Specifications ..................................................................................................40<br />

Appendix C. Warranty and Ordering Information ...........................................................................41


Table of Contents<br />

Figures<br />

1. Glyco Doc Imaging System ............................................................................................................1<br />

2. Installing the Camera Board in the Computer................................................................................3<br />

3. Glyco Doc Imaging System Cable Connections ............................................................................4<br />

4. Imager Power and Filter Indicators ................................................................................................5<br />

5. Glyco Doc Icon ...............................................................................................................................7<br />

6. Test Plate........................................................................................................................................8<br />

7. Image Display, Control, and Status Windows ................................................................................11<br />

8. SE1000 Camera Control Panel ......................................................................................................18<br />

9. Imaging System Functional Diagram .............................................................................................28<br />

10. Inside the Imager............................................................................................................................30<br />

Tables<br />

1. Preparing Gels for Imaging ............................................................................................................13<br />

2. Typical Exposure Strategies...........................................................................................................19<br />

3. Imager Troubleshooting..................................................................................................................34


Safety<br />

SAFETY<br />

!<br />

Warning<br />

Disconnect the Glyco Doc Imager before servicing. Refer servicing to <strong>Bio</strong>-<strong>Rad</strong><br />

service personnel.<br />

This Glyco Doc Imager is certified to meet the I.E.C. 1010* safety standard. Certified products are safe to<br />

use when operated in accordance with the instruction manual. This safety certification does not extend to<br />

other equipment or accessories not I.E.C. 1010 certified, even when connected to this Glyco Doc Imager.<br />

This instrument should not be modified or altered in any way. Alteration of this instrument will void the manufacturer’s<br />

warranty and create a potential safety hazard for the user.<br />

<strong>Bio</strong>-<strong>Rad</strong> is not responsible for any injury or damage caused by the use of this instrument for purposes other<br />

than for which it is intended or by modifications of the instrument not performed by <strong>Bio</strong>-<strong>Rad</strong> or an authorized<br />

agent.<br />

Important: This product conforms to the class A standards for Electromagnetic Emissions, intended for laboratory<br />

equipment applications. It is possible that emissions from this product may interfere with some sensitive<br />

appliances when placed nearby or on the same circuit as those appliances. The user should be aware<br />

of this potential and take appropriate measures to avoid interference.<br />

* I.E.C. 1010 is an internationally accepted electrical safety standard for laboratory instruments.


Installation and Startup<br />

1.0 INSTALLATION AND STARTUP<br />

The Glyco Doc Imaging System simplifies analysis of small format carbohydrate electrophoresis gels. Its<br />

primary components are the Imager, the Glyco Doc analytical software, and a host computer. The Imager is<br />

suitable for both fluorescent and visible dye applications. The Glyco Doc analytical software features automatic<br />

acquisition and analysis of fluorescent labeled oligosaccharide and monosaccharide gels, as well as<br />

DNA and Protein gels and gels stained with visible dyes.<br />

Figure 1. Glyco Doc Imaging System<br />

The Glyco Doc Imaging System provides the following features:<br />

• Integrated, high resolution CCD camera for gel imaging<br />

• Emission Filter switch, located on the Imager’s back panel, allows easy selection between Glyco<br />

Doc 500 nm and 600 nm imaging<br />

• Automatic exposure<br />

• Automatic band detection<br />

• Automatic quantitation and migration analysis<br />

• Support for numerous graphic file formats, including .tiff and .wmf<br />

• Export data and text for universal spreadsheet and word processing use<br />

• Report generator, for printing image display, band data, and densitometer readings<br />

1


Installation and Startup<br />

1.1 UNPACKING<br />

Carefully inspect the shipping cartons for any damage which may have occurred in shipping. Severe damage<br />

to a carton may indicate damage to its contents. If you suspect damage to the contents may have<br />

occurred, immediately file a claim with the carrier in accordance with their instructions before contacting <strong>Bio</strong>-<br />

<strong>Rad</strong> Laboratories.<br />

Open each of the shipping cartons and lift each component out of its packing. Check the contents against<br />

the supplied packing list. Inspect each component for external damage. If any part is missing or damaged,<br />

contact <strong>Bio</strong>-<strong>Rad</strong> Laboratories immediately.<br />

1.2 SET-UP THE COMPUTER AND IMAGER<br />

The setup procedure is organized into the following sections:<br />

• Section 1.2.1 is for those who purchased a system without a computer. This section discusses how<br />

to install the imager circuit board in your computer.<br />

• Section 1.2.2 is for all users. This section discusses how to set up all components in the system.<br />

1.2.1 Installing the Camera Circuit Board in a Computer<br />

Your host computer must be a standard IBM compatible PC, configured with the following minimum hardware<br />

and software:<br />

• 386 computer with math co-processor, running at 20 MHz or better, with at least 1 empty 16-bit<br />

expansion bus slot<br />

• 4 megabytes of RAM<br />

• VGA Display (640 by 480 resolution, 256 colors or shades of gray)<br />

• MS Windows and a compatible pointing device, such as a mouse<br />

In addition, because each Image file is approximately one half megabyte, the following storage devices are<br />

recommended.<br />

• A large-capacity hard disk (200MB or greater).<br />

• An optical disk drive. Although Images can be stored on floppy disks, an optical disk drive provides<br />

much greater storage capacity and provides high reliability. We recommend each user have their<br />

own removable optical disk to be stored in the same manner as other valuable media.<br />

!<br />

Caution/Warning<br />

Turn off and unplug the computer prior to removing its cover.<br />

2


Installation and Startup<br />

To install the camera board, follow the procedure below.<br />

1. Turn off your computer and unplug the power cord.<br />

This procedure assumes the computer was previously set up and used for other applications.<br />

2. Turn off all connected peripheral devices.<br />

Connected devices include printers, monitors, etc.<br />

3. Remove the cover from your computer.<br />

Refer to the documentation supplied with your computer system.<br />

4. Discharge static electricity.<br />

Make sure the computer is turned off and unplugged! Discharge static electricity by touching, with the<br />

back of your hand, a grounded metal surface such as the metal frame of the computer.<br />

5. Locate any unused 16-bit bus expansion slot inside your computer<br />

Expansion slots almost always run from the front of the computer to the back. A 16-bit slot is divided<br />

into two sections. (See Figure 2.)<br />

6. Remove the slot cover, which is located on the back of the computer.<br />

Removing the small slot cover on the back of the computer will allow you to attach a cable to the<br />

Camera board.<br />

Figure 2. Installing the Camera Board in the Computer<br />

3


Installation and Startup<br />

7. Place the Camera board in the slot<br />

The connector on the Camera board must face the back of the computer. Insert the board, pressing<br />

firmly so that it slips into place.<br />

8. Re-mount the screw removed in Step 6.<br />

This screw will hold the board in place.<br />

9. Re-mount the computer cover and its screws.<br />

Proceed to section 1.2.2, Setting up the System.<br />

1.2.2 Setting Up the System<br />

To set up the computer, you will need to make the connections shown below. Be sure to remove the small<br />

metal antistatic plug from the Imager’s computer cable connector. This plug protects the connector during<br />

shipping. Remove and save it for storage or shipment.<br />

Each cable is designed to fit a specific connector; where there is some doubt, note the number and configuration<br />

of the pins in the connector. Refer to Figure 3 and to the computer vendor’s documentation that was<br />

shipped with the computer. If you are using any peripheral computer devices with your system, refer to that<br />

device’s separate documentation for connecting to the computer.<br />

MONITOR<br />

IMAGER<br />

COMPUTER<br />

PRINTER<br />

KEYBOARD<br />

MOUSE<br />

Figure 3. Glyco Doc Imaging System Cable Connections<br />

4


Installation and Startup<br />

To set-up the Imager, you need a Phillips head screw driver. Refer to Figure 4.<br />

1. Locate the Filter Lock Screw on the bottom of the Imager and turn it counter clockwise, two complete<br />

turns.<br />

The Filter Lock Screw is encircled by an orange marker on the bottom of the Imager. The Filter is<br />

locked in place for shipping.<br />

2. Test to see that the filter is now free by twisting the Filter Switch.<br />

The Filter Switch is the black plastic knob on the back of the Imager. Looking from the back of the<br />

Imager, switch positions are 2 o’clock for 600 nm (Ethidium Bromide) and 10 o’clock for 500 nm (ANTS)<br />

labeled gels. Leave the switch at the 10 o’clock position. When the Imager is turned on, the selected<br />

filter will be indicated in the Imager’s front panel.<br />

Check for proper cabling and general system performance:<br />

1. Turn on the Imager.<br />

2. Turn on the computer and any peripheral devices.<br />

3. Check the Imager’s front panel to see that the Power and Filter Lights are lit.<br />

Rotate the Filter Switch to verify both Filter LEDs are working properly. See Figure 4 for the location of<br />

the Filter Switch.<br />

POWER LED<br />

500 nm FILTER LED<br />

600 nm FILTER LED<br />

BUSY LED<br />

ON/OFF<br />

SWITCH<br />

POWER<br />

OUTLET<br />

CONNECTOR<br />

TO COMPUTER<br />

FILTER<br />

SWITCH<br />

FRONT<br />

VIEW<br />

FILTER<br />

LOCK SCREW<br />

(LOCATED UNDER UNIT)<br />

REAR<br />

VIEW<br />

Figure 4. Imager Power and Filter Indicators<br />

5


Installation and Startup<br />

1.2.3 Installing the Glyco Doc Software<br />

Note<br />

If your purchase included a computer, proceed to section 1.3. Your computer<br />

was shipped with the Glyco Doc software installed.<br />

To install the Glyco Doc software, you need the following items:<br />

• 1 Glyco Doc setup disk<br />

• 1 Glyco Doc image disk<br />

To install the Glyco Doc software, follow the procedure below.<br />

1. Insert the Glyco Doc Setup disk into an available floppy drive.<br />

The computer must be turned on and display the DOS prompt or MS Windows startup screen. You may<br />

place the disk in any available 3.5” floppy disk drive. Typically, this is the a: drive. If you use the b:<br />

drive, substitute b: for a: in the following steps.<br />

2. Start the Install software.<br />

If the DOS prompt is displayed, type:<br />

win a:\setup.<br />

If the MS Windows Program Manager is displayed, select Run... from the File menu and then type:<br />

a:\setup<br />

3. Select the desired directory name where the Glyco Doc software files are to be placed.<br />

The installation routine will prompt for a directory name and location. The default is<br />

c:\glycodoc<br />

If a different directory is desired, enter that name instead. If you are re-installing the Glyco Doc software,<br />

the installation routine will overwrite any existing Glyco Doc files in this directory.<br />

4. When prompted, insert the Glyco Doc Image Disk into drive a: .<br />

The installation software will require a data subdirectory where image files will be stored. This will complete<br />

the installation.<br />

Note<br />

If you will not be using a large capacity removable media such as an optical<br />

disk drive, each user or project should have a subdirectory within the Glyco Doc<br />

directory. For example, Bob’s files might be stored in c:\glycodoc\bob.<br />

6


Installation and Startup<br />

1.3 VERIFY <strong>SYSTEM</strong> OPERATION<br />

This section provides a general description of the use of the Glyco Doc system.<br />

1.3.1 Starting the Glyco Doc Software<br />

When you power-up the computer, monitor, and Imager, a series of self-test and configuration status messages<br />

is displayed, followed by the C:\>, which is the indication that the system is ready.<br />

Start the Windows 3.1 interface software (by typing win), and then double click on the Glyco Doc icon to<br />

launch the Glyco Doc application. (Refer to the MS Windows User’s Guide for complete information on the<br />

use of MS-Windows 3.1.)<br />

Glyco Doc<br />

Glyco Doc<br />

Carbohydrate<br />

Analysis<br />

Figure 5. Glyco Doc Icon<br />

7


Installation and Startup<br />

1.3.2 Acquire an Image using the Test Plate<br />

To verify system operation, start by acquiring an image of the Test Plate.<br />

Note<br />

If you did not purchase a computer with the Glyco Doc system: Before doing<br />

any imaging, you must first create a Bright Field file. Refer to the chapter on<br />

Maintenance for discussion of the purpose of the Bright Field and the process<br />

for creating a Bright Field file<br />

IMAGE AREA SETUP PATTERN<br />

SE-1085<br />

Place cassette with the long plate down.<br />

Figure 6. Test Plate<br />

8


Installation and Startup<br />

To acquire an image using the text plate, follow the procedure below.<br />

1. Turn on the Imager and set the Filter Switch to 500 nm.<br />

The Power On/Off and Filter Switches are located on the rear of the Imager. The Filter Switch should<br />

be set to Carbohydrate, which uses a 500 nm filter.<br />

The LEDs on the front of the Imager indicate Power ON and the Filter setting.<br />

2. If you have not already done so, start the Glyco Doc software.<br />

Double-click on the Glyco Doc icon to start the software.<br />

3. Acquire an image.<br />

Select Acquire Options from the Image menu, and from the list that is displayed, select Select Image<br />

Rectangle.<br />

Follow the on-screen prompts. The first prompt instructs you to “Place the gel into the tray and slide the<br />

tray closed.” Place the Test Plate in the tray so that the silver strip along the edge faces into the Imager<br />

and the text printed on the test plate is not up-side-down. (See Figure 6.) Then select OK.<br />

The next prompt instructs you to select the area you want to image. To see the complete image, you<br />

may need to enlarge the display window. To do so, move the mouse so that the cursor is over the border<br />

of the image area; then hold down the left mouse button and move the mouse to enlarge the window.<br />

Then select OK.<br />

4. Acquire an image.<br />

Select Acquire from the Image menu, and from the list that is displayed, select Carbohydrate gel.<br />

5. From the SE1000 Camera Control window,<br />

a. Verify the Percent Saturation... radio button is selected, and it is set to 0.00.<br />

b. Verify the Normal Resolution radio button is selected.<br />

c. Click on the Acquire Image button in the SE1000 Camera Control window.<br />

Use the Up/Down arrows to change the setting.<br />

6. Follow the instructions presented.<br />

a. When prompted, place the Image Area Setup Pattern test plate into the gel holder.<br />

b. Follow the on-screen instructions.<br />

c. The acquired image is displayed. Enlarge the Image window, if necessary.<br />

7. Compare the resulting image with that of the test image printout found in the QC/Shipping packet.<br />

This printout was created using the same Image Area Setup Pattern test plate.<br />

If there are radical differences between the image you see and the test image printout provided with<br />

your imager, refer to the Imager Troubleshooting table in Chapter 4 to determine the problem and the<br />

appropriate corrective action.<br />

9


Installation and Startup<br />

1.4 IMAGE ANALYSIS USING THE IMAGE PROVIDED WITH YOUR <strong>SYSTEM</strong><br />

The key steps to analyzing an image are:<br />

• Open the image file (Use the image file demo1.RAW)<br />

• Adjust the image display<br />

• Find bands<br />

• Perform a standard lane analysis<br />

• Save the data file<br />

To analyze the sample image provided with your system, follow the procedure below.<br />

1. Open an image file.<br />

a. Select the Open command from the Files menu.<br />

b. From the Open File dialog box, select the image file demo1.RAW that has been provided.<br />

c. Press the OK button to open the file.<br />

2. Adjust the Image display’s Contrast and Brightness.<br />

The Image Histogram, shown in Figure 7, offers the following controls:<br />

• Contrast<br />

• Brightness<br />

• Autoscale, for automatically setting Contrast and Brightness.<br />

3. Locate bands using Glyco Doc’s automatic band location functions.<br />

These functions find all bands and mark with a + those bands whose data is displayed in the Band<br />

Tables.<br />

a. From the Bands pull-down menu, select either of the following:<br />

• To locate bands in a region, create a Marking Rectangle. Then, select Find Bands In Marked<br />

Rectangle.<br />

When defining a region, do not “crowd” the boundaries of a lane; include a small bit of the<br />

region beyond the band’s edge.<br />

• To find bands in the entire image, select Find Bands In Complete Image.<br />

b. Verify the number of lanes in the image area.<br />

Glyco <strong>DOC</strong> displays the number of lanes it has identified; if this number is not correct, enter the correct<br />

number.<br />

Note: When imaging the complete gel, not all lanes may be displayed; to see more lanes, pull the<br />

image window’s border to the right.<br />

c. Bands selected for analysis are automatically indicated by a + marker.<br />

To view all detected bands, select Show Band Outlines from the Options menu.<br />

4. Increase/decrease the number of bands marked for analysis.<br />

a. From the Bands menu, select Find More Find Fewer bands.<br />

b. Using the Find More or Less Bands window, point and click on the appropriate scroll bar arrow to<br />

quickly mark or unmark bands within the entire image.<br />

c. Press OK.<br />

10


Installation and Startup<br />

<strong>Bio</strong>-<strong>Rad</strong> Glyco Doc Analytical Software<br />

Files Image Bands Options Window Help<br />

1 2 3 4 5 6 7<br />

Mouse<br />

x=200 y=3<br />

Grey=5451<br />

Zoom<br />

Image Histogram<br />

Quan<br />

0-<br />

AutoScale<br />

Contrast<br />

Brightness<br />

Range: 5376 to 18175<br />

Figure 7. Image Display, Control, and Status Windows<br />

5. Selectively insert and mark a band or remove a marker from a band of no interest:<br />

• To remove a band’s marker, click on the + marker and select Delete in the Bands menu.<br />

When you point and click on a marker, it is highlighted in green.<br />

• To remove the markers from all bands in a selected lane, first select the lane by clicking on its Lane<br />

number. Then, click and drag the mouse to the ASingle Lane command in the Delete drop-down<br />

menu of Bands.<br />

• To insert and mark a band, select Insert Band from the Bands menu. The special cursor allows<br />

you to draw a band outline around the band.<br />

6. Perform analysis of the Standard Lane.<br />

a. From the Bands menu, select Standard Lane Analysis and specify the type of gel.<br />

b. Selecting Oligosaccharides displays the Click on Band window, which instructs you to move the<br />

cursor over the band which marks G4 in the standard lane and click on it. In the Quantification<br />

Value window, enter the number of pMols for the selected band.<br />

Selecting Monosaccharide, Protein, or DNA allows you to specify a standard by selecting from a list<br />

of standards or adding a new standard.<br />

11


Installation and Startup<br />

7. Analyze band data as presented in the Band Tables. The following tables are available:<br />

Band Location<br />

Description<br />

Band Location | Total Gray<br />

Rf<br />

Total Gray<br />

Degree of Polymerization<br />

Peak Gray<br />

Base Pairs<br />

Percent Luminance of Selected Bands<br />

Molecular Weight<br />

Quantity<br />

8. Perform other band analysis operations as desired.<br />

Each row in a band table displays bands which the software has determined are associated based on<br />

their location in the image (as measured in Y-axis pixel counts). In the image, associated bands are displayed<br />

by selecting a band; the crosshair in the selected band is displayed in green, and, when you<br />

select Show Row associations from the Options menu, the crosshairs for associated bands are displayed<br />

in blue.<br />

To change the sensitivity of the setting, select Associations... from the Bands menu and change the<br />

migration distance. The software then automatically re-defines all band associations.<br />

To modify band associations one band at a time, either:<br />

• Click on the band in the image and, while holding down the left mouse button, move the cursor to<br />

promote/demote the band to the next association level.<br />

• Click on the band value in any band table and, while holding down the left mouse button, move the<br />

data to the desired location.<br />

9. Save your work.<br />

Glyco Doc allows you to save both image and image data using the Windows 3.1 interface. When saving<br />

files, use the appropriate file extension, as shown below:<br />

Image file .RAW<br />

Band data file .BDF<br />

To save a file, use either the Save or Save As... command in the Files menu.<br />

• To save a new file, select Save and specify the filename.<br />

• To save changes to an existing file, select Save and do not change the filename.<br />

• To save changes to an existing file and at the same time keep the original file, select Save As... and<br />

enter a new filename. This allows you to save your work without overwriting the original file.<br />

10. Print reports, if you have a printer connected to the computer.<br />

The following printouts are available by selecting Print from the File menu:<br />

• Band Data Reports...<br />

• Densitometer Reports - The Densitometer report can be scaled in two ways:<br />

- Normalized to each Lane<br />

- Normalized to complete Image<br />

• Main Image Window<br />

• Screen<br />

12


Installation and Startup<br />

1.5 PREPARING FOR <strong>GEL</strong> <strong>IMAGING</strong><br />

This section discusses the preparation of gels prior to imaging. The table below provides some examples of<br />

how gels must be prepared before they are placed in the Imager tray.<br />

Table 1.<br />

Preparing Gels for Imaging<br />

FLUORESCENT <strong>GEL</strong>S<br />

CARBOHYDRATE <strong>GEL</strong>S<br />

(TAGGED WITH ANTS)<br />

<strong>GEL</strong> IN CASSETTE<br />

DNA <strong>GEL</strong>S (STAINED WITH<br />

ETHIDIUM BROMIDE)<br />

<strong>GEL</strong><br />

GLASS PLATE<br />

VISIBLE STAINED <strong>GEL</strong>S<br />

(USE PROTEIN SETTINGS)<br />

DNA <strong>GEL</strong>S<br />

( WITH SILVER STAIN)<br />

FLUORESCENT PLATE<br />

<strong>GEL</strong><br />

GLASS PLATE<br />

PROTEIN <strong>GEL</strong>S<br />

(STAINED WITH VISIBLE DYE)<br />

FLUORESCENT PLATE<br />

<strong>GEL</strong><br />

GLASS PLATE<br />

When preparing DNA and protein gels, follow the guidelines described below:<br />

• Inspect the glass plate for scratches and cleanliness.<br />

A glass plate is required with DNA and Protein gels. Hold each plate up to a light source while varying<br />

your viewing angle. If there are scratches, do not use the plate. Clean off any pieces of gel, fingerprints,<br />

liquids, or lint which will appear as artifacts in the image.<br />

• Place the gel onto the glass plate.<br />

Gently slide the gel onto the glass plate. Align the gel so that it is square on the plate. Remove bubbles<br />

from under the gel by running your finger across the top of the gel. Smooth out the wells so that the fluorescent<br />

plate will lay flat on top of the gel. Cut off any high ridge on the gel such as those found at the<br />

bottom of the gel.<br />

• Inspect the fluorescent plates for scratches and cleanliness.<br />

The fluorescent plate is required with Protein gels stained with visible dyes and DNA gels with silver<br />

stain. The purpose of the fluorescent bright field plate is to provide a visible (500 nm) light source for<br />

imaging. The UV light excites the fluorophore doped bright field plate wherever there isn’t a stained<br />

band, and it emits a negative image of visible light directly into the camera.<br />

If the fluorescent plate still has a protective coating, remove it while minimizing contact with its surface.<br />

Starting at one end of the gel, gently lower the plate, touching only one end of the plate down first.<br />

Continue to slowly lower the plate onto the gel, pushing out air bubbles as the plate is laid down.<br />

13


Installation and Startup<br />

1.6 EXAMPLE OF A STANDARD LANE ANALYSIS<br />

In the oligosaccharide sample image (demo1.raw) provided with your Glyco Doc system, there must be at<br />

least nine Band Markers in the oligosaccharide standard lane or the analysis will fail and the error dialog box<br />

is displayed. (More than nine markers may be present, as there are more than nine bands in the glucose<br />

ladder standard.) Standard bands G4 through G12 must have Band Markers. G4 is not as bright as the<br />

other bands in the lane and can be identified by this characteristic. It should have a marker since it is a key<br />

reference band in the oligosaccharide standard.<br />

If the number of bands does not agree with the set standard, then the analysis fails and the error dialog box<br />

appears.<br />

A procedure for a standard lane analysis is provided below.<br />

1. Remove any erroneous band markers from the standard lane.<br />

Shadow bands or blotches in the image are sometimes incorrectly marked during automatic band finding.<br />

Only the main bands in a standard lane should have band markers.<br />

a. Click on the unwanted band’s marker. The selected marker is displayed in green.<br />

b. Select Delete from the Bands menu or use the Delete key.<br />

2. Select a standard.<br />

a. Select Standard Lane Analysis from the Bands menu.<br />

b. In the Select a Standard dialog box, select Oligosaccharides.<br />

3. Select a band from the standard lane to serve as a reference for the lane.<br />

1. From the Click On Bands dialog box, indicate the band to be selected.<br />

2. Find the corresponding band in the image, and mark it by clicking on it. Then, press the OK button.<br />

For Oligosaccharides, the G4 band identifies the standard lane. (G4 is in the lower half of the standard<br />

lane and appears approximately one quarter as bright as the other bands in the lane.) It is used to indicate<br />

a Degree of Polymerization (DP) of 4, and a quantity of 50 pMols. In the default settings G4 is<br />

given an Rf=1, G12 is given an Rf=0.<br />

4. Enter the band’s quantification value.<br />

In the Quantification Value window, enter the number of pMols of sugar for the band selected in step 3.<br />

Then press OK.<br />

For purposes of this demonstration, enter: Oligosaccharide sample: G4 = 50.<br />

A standard lane’s designator is the letter “s”, rather than a number.<br />

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Software Reference<br />

2.0 SOFTWARE REFERENCE<br />

This chapter discusses each of the software functions. The chapter is organized according to the software’s<br />

menu structure.<br />

2.1 FILES MENU<br />

Each of the Files drop down menu’s selections is discussed below.<br />

Open...<br />

Displays the Open File window, which lists all the files available to Glyco Doc and their location on the computer’s<br />

hard disk.<br />

Close<br />

Closes the currently loaded file. If a new image is currently displayed, or the band data has been changed,<br />

the system prompts with a message asking if you first want to save your work. Selecting Yes displays the<br />

Save Files window, which is discussed below.<br />

Save<br />

Allows you to save both the image and the image’s data. Select Save when saving a new file or saving<br />

changes to an existing file. This displays the Save Files window. When saving a new file, use the appropriate<br />

file extension, as shown below:<br />

Image file .RAW In the Save Files window, point and click on the Gel Image<br />

(.RAW) checkbox.<br />

Band data file .BDF In the Save Files window, point and click on the Band Data<br />

(.BDF) checkbox.<br />

The Save Files window also indicates whether the disk contains a previous version of either file. To prevent<br />

writing over a previous version of the file, press New Name. This displays the Enter Base Filename To<br />

Save As window (a Save As window), which allows you to give the file a new name.<br />

Since an image file is typically 500,000 bytes in size, save files only as necessary; their large size quickly<br />

consumes disk space. The 230 MB rewritable optical disks provided with some systems are intended for<br />

the storage of image files.<br />

Save As...<br />

Selecting Save As... allows you to enter a new filename, so that you can save your work without overwriting<br />

the original file.<br />

Print<br />

Glyco Doc allows you to print band data and densitometer reports, as well as printing the image and all the<br />

windows displayed on the computer screen.<br />

Band Data Reports...<br />

Includes the data from any of the band tables.<br />

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Software Reference<br />

Densitometer Reports<br />

A printout is generated for each lane, with the Densitometer positioned next to its corresponding lane.<br />

All quantitative and migration data for that lane is included. The Densitometer report can be scaled in<br />

two ways:<br />

- Normalized to each Lane - This report scales the densitometer output so that the brightest peak in<br />

each lane corresponds to the top of the densitometer scale for that lane’s output.<br />

- Normalized to complete Image - This report scales the densitometer output so that the brightest<br />

peak in the gel corresponds to the top of the densitometer scale.<br />

Main Image Window<br />

Prints out the image using the current contrast and brightness settings. The printout does not include<br />

any band markers or highlighted pixels.<br />

Screen<br />

All windows appearing on the screen are printed. Images printed out by this command use the current<br />

brightness, contrast, and reverse palette settings.<br />

Print Setup<br />

This is an MS-Windows function that lets you specify the printer and print quality.<br />

Import Image<br />

This function allows you to import images from other applications of from other imaging systems such as<br />

<strong>Bio</strong>-<strong>Rad</strong>’s Gel Doc systems.<br />

Export<br />

Glyco Doc images and data can be exported to other applications.<br />

Band Data to Spreadsheet...<br />

Band data is exported in a generic, tab separated, text spreadsheet format. The gray values shown in<br />

the Pixel Values window can also be exported in a generic, tab separated, text spreadsheet format. To<br />

export gray values in a marked region, select the Export command’s Marked Rect. Values to<br />

Spreadsheet...option.<br />

Marked Rectangle Values to Spreadsheet...<br />

This function allows you to export the Marked Rectangle gray values to a spreadsheet for analysis.<br />

Copy Marked Rectangle to Clipboard<br />

This function allows you to copy the Image area contained within the Marked Rectangle. That portion of<br />

the image then can be pasted into a paint application.<br />

Copy Band Data to Clipboard<br />

This allows you to export by using MS Windows Clipboard.<br />

16


Software Reference<br />

Image File<br />

To export an image from Glyco Doc, so that it can be incorporated into a report or document created in<br />

a different application. In the Save As window that is displayed,<br />

1. Use the File Name field to change the displayed filename’s extension to .RAW, which is required of<br />

all Image files.<br />

2. From the list of Image files that is displayed, select the file you want to save.<br />

3. From the Save File as Type field, select the file type supported by your software application. The<br />

following file types are available, along with some of the operating systems and software applications<br />

which support each file type. (If you are not sure which type to select, refer to your software<br />

application’s technical documentation.)<br />

• TIFF - Tagged Image File Format • WMF - Windows Meta File<br />

• PCX - ZSoft and Paint Brush • DCX - Multipage pcx<br />

• BMP - MS Windows and OS/2 Bitmap • EPS - Postscript<br />

• T&arga - Truevision • Jpeg - JPEG<br />

• GIF - CompuServe • PCT - Macintosh Pict<br />

• Wpg - Wordperfect<br />

Exit<br />

This selection quits the Glyco Doc application.<br />

17


Software Reference<br />

2.2 IMAGE MENU<br />

To obtain the best image from which both image and band data files are derived, the Image menu offers a<br />

number of setup controls and options.<br />

Acquire<br />

This selection initiates image acquisition. It requires you to specify the type of gel to be imaged:<br />

Carbohydrate, DNA, or Protein, before bringing up the SE1000 Camera Control window.<br />

SE 1000 Camera Control<br />

Acquire using<br />

Percent Saturation... 0.00<br />

(between 0.0 and 5.0)<br />

Exposure time... 1.00<br />

(0.1 secs to 10.0 sec:)<br />

Acquire using<br />

SE 1000 Camera Control<br />

Normal Resolution<br />

High Resolution<br />

(Takes twice as long)<br />

Background Percent of Saturation<br />

(between 5% and 75%)<br />

25<br />

Acquire Image<br />

Cancel<br />

Acquire Image<br />

Cancel<br />

SE 1000 Camera Control for Carbohydrate and DNA Gels<br />

SE 1000 Camera Control for Protein Gels<br />

Figure 8. SE1000 Camera Control Panel<br />

Percent Saturation and Exposure time<br />

Exposure Time is the length of time that the camera requires to acquire an image. Percent Saturation<br />

sets a limit on the percentage of detectors and corresponding pixels that are saturated within a region.<br />

If this is the first time a gel is being imaged, start by setting the Percent Saturation. Refer to Table 2 for<br />

examples of typical Percent Saturation settings. After you have acquired the image, you may see areas<br />

of yellow. This is an indication that saturation of the corresponding CCD detectors has occurred.<br />

Saturation occurs when the individual cells or pixels in the CCD camera can no longer quantify additional<br />

light (they have reached their capacity). You should see little or no yellow in bands to be quantified or<br />

in bands that are being used as a quantification standard. Saturation is a problem only if it affects the<br />

imaging of bands which are to be quantified or are to serve as the basis of measurement. Otherwise,<br />

bands which are not of interest may show signs of saturation without affecting the analysis data.<br />

Continued saturation can lead to “blooming,” an effect in which the CCD detectors surrounding a saturated<br />

detector in turn become saturated; when looking at the image, you can see “blooming” where the<br />

yellow indicating saturation seems to spill over beyond the boundary of the band.<br />

If any of the bands of interest was saturated, note the exposure time used to acquire the saturated<br />

image. Use this time as a starting point for a time exposure. Reduce the exposure time to reduce the<br />

saturation. An optimal exposure can take several iterations.<br />

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Software Reference<br />

The Exposure Time is influenced by how intensely the fluorophore is emitting light. This, in turn, is proportional<br />

to the concentration of the substance containing the fluorophore in the gel; greater intensity<br />

means shorter exposure. The Exposure time... setting should prevent overexposure of bands of interest;<br />

saturation is acceptable only if it is indicated for bands that will not be quantized.<br />

To see the results of overexposure, select HiLite Saturated Pixels from the Options menu. This selection<br />

functions as a toggle: If a checkmark appears next to your selection, saturated areas are displayed<br />

in yellow in the image and the amount of saturation can be read in the Pixel Table. To turn off the display<br />

of saturated areas, again select HiLite Saturated Pixels.<br />

To detect underexposure, move the Marking Cursor over the centers of the bands of interest. As you<br />

move the cursor watch the Mouse Table. The Gray pixel values should range between 50,000 and<br />

60,000. If the center of the band shows a value below 50,000, the image may be considered underexposed.<br />

If the image shows lanes of differing intensities, adjust the SE1000 Camera Control window’s Percent<br />

Saturation... to limit the percentage of detectors and corresponding pixels that are saturated within a<br />

region. For example, if your band(s) of interest were averaging a 12,000 pixel value at a .8 second<br />

exposure time, you might want to take a second exposure with the exposure time set to 1.6.<br />

The table below provides guidelines for typical gels, and is intended as a starting point for imaging.<br />

Table 2.<br />

Typical Exposure Strategies<br />

Image Type Percent Saturation Exposure Time<br />

Oligosaccharide 0 - 5 –<br />

Monosaccharide - 6 seconds<br />

Protein 25 –<br />

DNA 0 - 5 –<br />

Note that prolonged or repeated exposure of the gel to UV light can lead to bleaching, which is the result<br />

of exposing a fluorescently tagged gel to the UV light source for a period which causes the fluorophore<br />

to diminish its light output.<br />

Image Resolution<br />

The image may be acquired for display at Normal Resolution or High Resolution. To capture the image<br />

and present it as data, a series of exposures is taken whenever the camera is triggered. The data is<br />

averaged to create the final pixel values which are used for display and analysis. Selecting High<br />

Resolution doubles the number of exposures that are taken. This yields a higher image quality, but risks<br />

bleaching the gel. For this reason, Normal resolution is recommended.<br />

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Software Reference<br />

Acquire Options<br />

These selections must be made before you acquire the image.<br />

Select Image Rectangle<br />

After you acquire the first image of a gel, you may be able to improve the quality of subsequent images<br />

by drawing an Image Rectangle over your area of interest. (The sensitivity of the camera becomes<br />

greater as you reduce the extremes of intensity in the imaging area.) To mark a region using the Image<br />

Rectangle,<br />

1. Place the cursor where you want the first corner of the Image Rectangle. Note that the cursor<br />

becomes the Marking Cursor when it is in the image area of the screen.<br />

2. Click and drag the cursor to draw a rectangle over the desired area.<br />

3. Release the mouse button when the desired region is enclosed.<br />

4. Notice that the histogram information changes to reflect the marked region.<br />

A previously drawn Marking Rectangle remains in effect until a new one is defined.<br />

Image Size<br />

Image size is selected in the Image Display Options window. The image may be acquired for display in<br />

one of the following sizes:<br />

• Medium format. This is the most common since it yields an image which is easily viewed and a<br />

manageable file size.<br />

• Small format. This results in a smaller image on the screen and a smaller image file. (This is<br />

accomplished by the software, which averages the values of four pixels and outputs that averaged<br />

value for a single displayed pixel.) However, the image data available for analysis in the Band<br />

Tables is the same as with other formats.<br />

• Large format. This provides the largest image display, and results in a larger image file size.<br />

However, the data available in the Band Tables is the same as that for the other formats.<br />

Make New Bright Field File<br />

The Bright Field File serves as a reference image for the subsequent imaging of gels. It allows the software<br />

to correct for any imperfections in the optics on the CCD camers. Carbohydrates and Proteins<br />

share the same Bright Field file; DNA imaging requires its own Bright Field file. If any components of<br />

the optical path (such as the lenses, filters, UV light, mirror and camera) are cleaned, moved, replaced,<br />

or altered in some manner, a new Bright Field file must be created. For detailed discussion of the Bright<br />

Field file, refer to Maintenance and Troubleshooting.<br />

Image Information<br />

To enter a description or record notes for any particular image file. In the window that is displayed, enter the<br />

following:<br />

Description: Enter a short description for the information you want to save.<br />

Edit Notes: Click on this button and in the window that is displayed, you can enter and save notes on<br />

the Image.<br />

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Software Reference<br />

2.3 BANDS MENU<br />

Glyco Doc identifies bands by looking for sudden drop-offs in intensity. Glyco Doc then groups bands<br />

according to the following definitions:<br />

• Lane: Glyco Doc defines a Lane as all bands in the same column.<br />

• Row: Glyco Doc assigns a Row Association to all bands appearing at approximately the same distance<br />

from the top of the image.<br />

Glyco Doc also allows you to insert a band not automatically located by the software and to change a band’s<br />

row association.<br />

Image data for all bands is displayed in the Band Tables. For a band’s image data to be displayed, that<br />

band must be marked. You can use the software to automatically mark bands (Find Bands), or you can<br />

selectively insert or delete a band’s mark. Band tables provide the most detailed representation of image<br />

data. Band tables are organized into lanes and rows, which correspond to the locations of marked bands.<br />

Find Bands in Complete Image<br />

This function automatically locates and identifies all bands in the image and inserts band markers. A band<br />

must first be marked before its data is displayed in the Band Tables. Bands are marked based on size and<br />

intensity criteria.<br />

Find Bands in Marked Rectangle<br />

This selection differs from finding bands in the complete image only in that it allows you to specify your area<br />

of interest before band finding and band marking. To Find Bands in a Marked Rectangle, start by drawing a<br />

Marking Rectangle over your area of interest before selecting Find Bands in Marked Rectangle. When<br />

defining a region, do not “crowd” the boundaries of a lane.<br />

Prompt for Number of Lanes<br />

This is a toggle function which allows you to confirm the number of lanes that the software will label during<br />

the automated band finding routine. Because the number of lanes detected may not always be correct, you<br />

may want to select this function so that you can confirm - or correct - the number of lanes found.<br />

Number of Wells on Gel: Selecting this function allows you to confirm the number of wells on the gel. The<br />

number of wells may be different than the number of lanes.<br />

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Software Reference<br />

Insert Band<br />

This selection allows you to insert a band which may otherwise have been overlooked by the automatic<br />

band marking function. Selecting Insert Band causes the Outline cursor to be displayed. By pressing the<br />

left mouse button and dragging, you draw the band’s outline. When you release the left mouse button, the<br />

band’s marker appears and the band data is included in the band tables.<br />

Delete<br />

The three choices under this selection delete band data from the band tables. To return a band’s data to the<br />

band tables after a delete operation, you must use the band finding functions (Find Bands in Complete<br />

Image or Find Bands in Marked Rectangle) or use the Insert Band function.<br />

Band<br />

Use this function to delete a band’s data from the band tables. Click on the + marker to select the band<br />

(the mark is green when selected), and then select Delete - Band. Deleting a Band Marker does not<br />

remove that band’s outline, but it does delete that band’s data from the band data tables.<br />

ASingle Lane<br />

To remove all data for a selected lane from the Band Tables. Note that the Lanes are re-numbered.<br />

All Bands and Lanes<br />

Deletes all band and lane data from the band tables.<br />

Find More Find Fewer<br />

This function allows you to adjust the sensitivity of Glyco Doc’s automatic band marking software to automatically<br />

increase or decrease the number of marked bands. Note that it affects the number of bands<br />

marked, but not the number of bands found. From the Find More or Less Bands window, use the appropriate<br />

scroll bar arrow to quickly mark or unmark bands within the entire image.<br />

Standard Lane Analysis<br />

For standard lane analysis, each band must be associated with a standard band before it can be assigned a<br />

quantity. Glyco <strong>DOC</strong> supports the following standard lane analyses. An example of a standard lane analysis<br />

is provided at the end of the previous chapter.<br />

Oligosaccharide<br />

This uses the glucose ladder standard.<br />

Monosaccharide<br />

This uses the monosaccharide ladder standard.<br />

DNA<br />

This allows you to specify your desired standard.<br />

Protein<br />

This allows you to specify your desired standard.<br />

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Software Reference<br />

Set Quantitive Standard Band...<br />

This allows you to select a band other than the standard.<br />

Display Standard Plot on Log Scale<br />

This allows you to plot Size versus Rf for Protein and DNA gels.<br />

Band Information<br />

This is the data available for analysis and displayed in the band tables. (For a complete discussion of the<br />

Band Tables, refer to that section later in this chapter.) To display a band’s information, double click on its<br />

marker, or click once and select Band Information....<br />

Lane Information<br />

This contains a description of the lane. To display a lane’s information, double click the lane’s number, or<br />

click once and select Lane Information...<br />

• Lane Number: The lane number is displayed at the top of the image.<br />

• Number of Bands: The number of bands in the lane<br />

• Total pmols: The total of all bands in that lane.<br />

• Lane Description: Enter any description for the lane.<br />

Associations...<br />

During automatic band marking, bands in different lanes, having similar migration distances (as measured in<br />

Y-axis pixel counts), are automatically grouped into “Row Associations.” Band associations can be observed<br />

in band tables and in the image display.<br />

To change the sensitivity of this setting, select Associations... from the Bands menu. This changes the<br />

pixel count value used by the software to assign associations. The software then automatically re-defines all<br />

band associations.<br />

Band Associations can be manually adjusted one band at a time by clicking on the band value in any band<br />

table and, while holding down the left mouse button, moving the data to the desired location. Band associations<br />

can also be manually adjusted in the Image window by clicking and dragging the band to its new association.<br />

When a band is clicked on and dragged, a Linking Cursor appears. When you move it, the cursor<br />

changes to an Up or Down arrow until you release the mouse button.<br />

To remove a band from a band association, click on it and drag it from its row. If you drag to the top or bottom<br />

of the image, a new row is created in the band table, with the band placed in that row.<br />

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Software Reference<br />

2.4 OPTIONS MENU<br />

The Options menu provides a number of display selection, each of which is discussed below.<br />

Reverse Palette<br />

When selected, the displayed image is reversed, so that a negative image becomes positive and a positive<br />

image becomes negative.<br />

HiLite Saturated Pixels<br />

When selected, saturated pixels are displayed in yellow.<br />

Autoscale every image update<br />

When this is selected, the software automatically sets the Contrast and Brightness for the entire image or<br />

Image Rectangle. This is the equivalent of clicking the Auto Scale button in the Image Histogram.<br />

Hide Band Marks<br />

Allows you to hide the band markers without removing the band data from the band tables.<br />

Show Lane Center Line<br />

The band finding function includes the identification of lanes. Along with indicating Lanes by a number, you<br />

can also indicate lanes by a line drawn through their centers. Selecting a lane by clicking once on its number<br />

changes the lane’s center line to green.<br />

Show Row associations<br />

This allows you to turn on/off the display of band associations. Associated bands are displayed in blue.<br />

(See Associations... under the Band menu.)<br />

Show Band Outlines<br />

This function allows you to see each of the bands found by the software. This allows you to verify that all<br />

bands of interest have been identified. Glyco Doc identifies a band’s edge by determining where there is a<br />

rapid drop-off in intensity. Where a band of interest has not been detected, or two adjacent bands are identified<br />

by a single band outline, you can use the Insert Band function in the Band menu to selectively insert<br />

bands.<br />

24


Software Reference<br />

2.5 WINDOW MENU<br />

This menu allows you to specify what type of data you want to display.<br />

Band Table<br />

A band table is a resizable and scrollable window displaying data about each marked band. This selection<br />

is a toggle function for hiding or displaying band tables.<br />

The different band tables are available by clicking on the word “Mode” in any displayed band table. Note the<br />

following:<br />

• This is selected from the upper left corner of any band table. It is not a menu bar item.<br />

• Selecting a mode does not display that band table; you must also have Band Table selected.<br />

Band Location<br />

The location of a band is measured in pixels on an X/Y axis. This table displays the Y-axis value of the<br />

center of each marked band (as indicated by its cross-hair). Since the band is associated with a lane,<br />

its X-axis value is not important. The corrected value refers to the value assigned by the software after<br />

it has accounted for any curving up or down of the gel.<br />

Band Location/Total Gray<br />

The Band Location values are the corrected values from the Band Location table. The Total Gray value<br />

for a band is the total intensity for all pixels in that band.<br />

Total Gray<br />

These values are the same as those presented in the Band Location/Total Gray table.<br />

Peak Gray<br />

This is the largest gray value (up to a value of 65,000) in the selected band. (This value does not necessarily<br />

correspond to the center of the cross-hair.)<br />

Percent luminance of Selected Bands<br />

This presents the relative intensity (percentage of luminance) for each band in a selected lane. The<br />

total for all bands in a lane is 100%. (The data in this table is similar to that in the Total Gray band<br />

table, which presents the data as the total intensity for all pixels in a selected band.)<br />

Quantity<br />

This is the concentration (in picomols) of the marked bands.<br />

Description<br />

This is any text you might choose to enter. To enter text from the band table, double click on the desired<br />

band’s field or on the desired band in the image. Either method displays the Information about<br />

Selected Band window where you can enter the description.<br />

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Software Reference<br />

Rf<br />

Rf (Relative mobility) are determined by setting the center point of one band to have an Rf = 1 and that<br />

of another band to have an Rf = 0. All bands between those two points are given a value between 1<br />

and 0 based on a linear scale. Bands above Rf = 0 are given a negative value and those below Rf = 1<br />

are given a value greater than 1. These numbers are based on the same scale as the numbers in the 0<br />

to 1 range. For oligosaccharides, the Rf = 1 band is G4 and the Rf = 0 band is G12.<br />

Degree Polymerization (Dp)<br />

Degree of Polymerization is used during carbohydrate gel analysis to provide positional information for<br />

each band based on glycose units. This information is used by the software to establish band associations.<br />

Base Pair<br />

Base Pair is available when evaluating a DNA gel. This measurement provides migration data based on<br />

a base pair standard.<br />

Molecular Weight<br />

Molecular weight data is provided when evaluating Protein gels.<br />

Mouse Table<br />

This table shows the current location (measured in pixels along an X/Y axis) of the cursor in the image area,<br />

and the grayscale intensity of that pixel.<br />

Pixel Table<br />

Shows the grayscale intensity of every pixel in the image. To view the intensity of a pixel, :<br />

• Click once on the value in the Pixel Table and watch that pixel flash in the image.<br />

• Click once on a pixel to display its value in the Pixel Table. Or, you can draw a marking rectangle to<br />

view the pixel values of any number of pixels.<br />

Densitometry Graph<br />

The Densitometer graphically represents changes in intensity across a gel image. Its modes of representation<br />

allow you to display different intensity levels within a lane, between lanes, or within a marked region.<br />

These modes are defined below. You can select a lane by clicking on the lane’s number. Select multiple<br />

lanes by clicking on successive lane numbers while holding down the key.<br />

Densitometry Modes<br />

The densitometer offers two modes:<br />

Show Marked Rectangle<br />

In this mode, the Densitometer displays a graph showing the average intensities in the marked region.<br />

The size of the graph will change to reflect the height and location of the region.<br />

26


Software Reference<br />

Show Selected Lanes<br />

This mode displays the intensity levels within a lane or compares between two or more lanes. Include<br />

the first lane by clicking on the number found at the top of the lane. Include additional lanes by holding<br />

down the key and clicking on each additional lane number. Each lane will have its fluorescent<br />

intensity plotted in the graph. Each lane number is displayed in a different color at the top of the image<br />

display and matches the color of the lane number in the densitometer graph .<br />

Histogram<br />

Once an image has been acquired, there are a number of adjustments that can be made using the Image<br />

Histogram to improve the quality of the display. Changing the display using the Image Histogram has NO<br />

EFFECT on image data<br />

The Image Histogram window provides the following controls and display information:<br />

• A curve showing the relationship between the number of pixels in the entire image or the Image<br />

Rectangle versus the intensity of that image area evaluated for 65,535 shades of gray.<br />

• AutoScale button: Pressing this button automatically sets the Contrast and Brightness for the entire<br />

image or the Image Rectangle. It does this by setting the histogram curve’s cursors (indicated by<br />

red, dashed vertical lines) to the region of greatest activity, which is much narrower than the full<br />

range of 0 to 65,535. This function will provide a good image for most gels; the Brightness and<br />

Contrast controls then can be used to improve the quality of the display.<br />

• Range: These numbers are the lower and upper limits of the boundary defined by the histogram<br />

cursors.<br />

• Contrast and Brightness: These can be controlled either by using the sliders or the vertical cursors<br />

(indicated by red, dashed lines) in the histogram curve. Use Contrast to change the distance<br />

between the cursors, as indicated by the difference between the upper and lower range values.<br />

(Note: The difference between the range values cannot be less than 256, since that would degrade<br />

the monitor’s performance.) Use Brightness to shift the cursors left or right.<br />

Click on and drag directly on a slider to increase or decrease a setting, or click on an arrow to move<br />

the box back and forth in incremental steps.<br />

Zoom Window<br />

This window allows you to zoom in on the image under an image rectangle. This function can be considered<br />

as a magnifying glass, where magnification is controlled by the size of the Zoom window. To zoom in,<br />

draw an image rectangle over the area of interest and then click on it. You can also zoom in by clicking and<br />

holding down the right mouse button; .<br />

27


Theory of Operation<br />

3.0 THEORY OF OPERATIONS<br />

During exposure, the Imager’s CCD (Charge Coupled Device) detector collects photons emitted by the fluorophore<br />

in a prepared gel. The detector is a semiconductor grid of approximately 180,000 cells for collecting<br />

these photons. Each cell in the grid can collect up to 65,535 charge units per exposure. If a cell receives<br />

more than 65,535 charge units, the excess charge units spill into the surrounding grid squares (in a phenomena<br />

called “blooming”), and that area of the gel is considered saturated. These values serve as the raw<br />

data for band analysis.<br />

Because computer monitors typically display a maximum of 256 shades of gray, the computer must scale<br />

each grid cell’s value to a value between 0 and 256, where 0 is absolute white, and 256 is absolute black.<br />

Each “pixel” (picture element) on the monitor is illuminated at a corresponding level of brightness. This<br />

value is a pixel’s grayscale value.<br />

UV SOURCE<br />

(350nm to 400 nm)<br />

EXCITATION FILTER<br />

(NOT USER-SELECTABLE)<br />

<strong>GEL</strong><br />

EMISSION FILTER<br />

(FILTER TYPE IS SELECTED<br />

USING THE SWITCH ON THE<br />

BACK OF THE IMAGER)<br />

CCD CAMERA/<br />

DETECTOR<br />

Figure 9. Imaging System Functional Diagram<br />

28


Maintenance and Troubleshooting<br />

4.0 MAINTENANCE AND TROUBLESHOOTING<br />

In this chapter, the following Imager hardware maintenance procedures are given:<br />

• Replacing the UV Light<br />

• Cleaning the Excitation Filter<br />

• Creating a New bright field<br />

A trouble-shooting chart for the Imager is provided in Section 4.5. For trouble-shooting computer problems,<br />

consult the appropriate computer vendor’s documentation. For difficulties with memory storage limitations,<br />

see Section 4.6.<br />

!<br />

Warning<br />

Disconnect the Glyco Doc Imager before servicing. Refer servicing to <strong>Bio</strong>-<strong>Rad</strong><br />

service personnel.<br />

4.1 REPLACING THE UV LIGHT<br />

The UV light should be replaced after 6 months of continuous use. For this procedure, you will need a<br />

Phillips head screw driver.<br />

1. Turn off the computer and the Imager.<br />

2. Unplug the power cord from the back of the Imager.<br />

3. Disconnect the Imager Interface Cable.<br />

4. Remove the Imager’s Top Cover.<br />

Slide out the gel holder to reveal two Phillips head screws obscured by the holder’s front panel. Four<br />

more Phillips head screws are located on the outer edges of the rear panel. Remove all six screws and<br />

then lift off the Top Cover. Save the screws.<br />

5. Detach the two Wire Bundles from the Reflector.<br />

Be careful not to bend or distort the Reflector.<br />

6. Remove the Reflector.<br />

Locate and remove the shiny tent-like reflector held in place by two thumb screws. Be careful not to<br />

bend or distort the reflector.<br />

At this time you may wish to inspect the Excitation Filter and clean if necessary.<br />

7. Remove the UV Tube.<br />

Grasp and twist the UV tube 90° to release and remove it from the holder. Applying outward pressure to<br />

the back of the Imager Chassis may help. When inserting a new tube, be sure to align it so that the<br />

manufactures label faces upward toward the top of the reflector.<br />

29


Maintenance and Troubleshooting<br />

8. Perform the previous steps in reverse to install the new tube and close-up the Imager.<br />

9. Create a new Bright Field file.<br />

Refer to Section 4.4.<br />

EXCITATION<br />

FILTER<br />

EMISSION<br />

FILTER<br />

Figure 10. Inside the Imager<br />

30


Maintenance and Troubleshooting<br />

4.2 CLEANING THE EXCITATION FILTER<br />

The Excitation Filter is located immediately above the gel holder and just below the UV lamp. It is a glass<br />

square approximately 3.75” and is a metallic brown color. Follow the procedure below to replace the<br />

Excitation Filter.<br />

Note<br />

Wear lint and powder free gloves while holding the filter.<br />

1. Turn off the Computer and the Imager.<br />

2. Unplug the power cord from the back of the Imager and disconnect the Imager Interface Cable.<br />

3. Remove the Imager’s Top Cover.<br />

Slide out the gel holder to reveal two Phillips head screws obscured by the holder’s front panel. Four<br />

more Phillips head screws are located on the outer edges of the rear panel. Remove all six screws and<br />

then lift off the Top Cover. Save the screws.<br />

4. Detach the two Wire Bundles from the Reflector.<br />

Be careful not to bend or distort the Reflector.<br />

5. Remove the Reflector.<br />

Locate and remove the shiny tent-like reflector held in place by two thumb screws. Be careful not to<br />

bend or distort the reflector. You may wish to inspect the Excitation Filter and clean if necessary.<br />

6. Remove the UV Tube.<br />

Grasp and twist the UV tube 90° to release and remove it from the holder. Applying outward pressure to<br />

the back of the imager chassis may help.<br />

7. Remove the Excitation Filter.<br />

Locate and remove the thumb screws which hold the filter in place. Then, grasp the filter by its edges<br />

and remove it from the Imager.<br />

8. Clean the Excitation Filter.<br />

Use dry oil free compressed gas to blow particulates off the filter.<br />

The filter at this time should be free of any residue. If not, clean with 50% alcohol in water which has<br />

been stored in glass only. Alcohol which has been stored in plastic will leave a fluorescent residue on<br />

the filter.<br />

9. Replace the Excitation Filter with the most blemish free side facing downwards.<br />

10. Insert the new tube and close up the Imager.<br />

11. Create a new Bright Field file to complete this procedure.<br />

Refer to Section 4.4.<br />

31


Maintenance and Troubleshooting<br />

4.3 PREPARING <strong>GEL</strong> PLATES<br />

The gel plates should be free of smudges, blotches, or any film on both sides. Clean off any pieces of acrylamide,<br />

fingerprints or lint which will appear as artifacts in the image. If the glass needs to be cleaned, we<br />

recommend using Kimwipes EX-L Lens Cleaning Station, sold as Kimwipes No. 34662, consisting of two<br />

boxes of Kimwipes EX-L delicate Task Wipers and 1 bottle of Kimwipes EX-L Lens Cleaning Solution. If not<br />

wiped completely dry, this solution leaves a film on the plate.<br />

4.4 CREATING A NEW BRIGHT FIELD<br />

Components of the optical path include the lenses, filters, UV light, mirror and camera. If any of these items<br />

are cleaned, moved, replaced, or altered in some manner, a new Bright Field file must be created. The purpose<br />

of the Bright Field File is to obtain a reference image using the current optical path.<br />

Changes to the optical path include:<br />

• Changing the UV light<br />

• Cleaning a Filter<br />

• Cleaning and realigning the mirror<br />

• Changing the Lens<br />

You will need the following materials:<br />

• 1 Clean, scratch free, Bright Field Plate<br />

• 1 Test Plate<br />

Note<br />

Handle the Bright Field Plate by its edges only. Once finished immediately<br />

place the plate in the supplied bag. Store the plate in a manner which will keep<br />

it free of scratches.<br />

To create a new bright field, follow the procedure below.<br />

1. Inspect the Bright Field Plate. (This is the green glass plate supplied with your system.) To create<br />

a new Bright Field file you will need a clean Bright Field Plate with no scratches.<br />

If the plate still has the protective coating, remove it while minimizing contact with its surface. Hold the<br />

plate up to a light source while varying your viewing angle. If scratches are seen, dispose of the plate.<br />

The plate should be free of smudges, blotches, or film . <strong>Bio</strong>-<strong>Rad</strong> recommends Kimwipes EX-L Lens<br />

Cleaning Station. Sold as Kimwipes No. 34662, it consists of two boxes of Kimwipes EX-L Delicate Task<br />

Wipers and 1 bottle of Kimwipes EX-L Lens Cleaning Solution. If not wiped completely this solution will<br />

leave a film on the plate.<br />

32


Maintenance and Troubleshooting<br />

2. From the Image menu’s Acquire Options, select Make New Bright Field File.<br />

Follow the on-screen instructions, directing you to insert the Bright Field Plate. After the Bright field<br />

image is acquired, remove the plate.<br />

3. Save the Bright Field file.<br />

After selecting OK at the prompt to save, the file is stored as se1000.brt.<br />

The Bright Field file is stored in the directory specified by the line shown below, written in the glyco.ini<br />

Glyco <strong>DOC</strong> initialization file:<br />

DefaultBrightFileDir=c:\glycodoc\data\<br />

The text to the right of the equals sign is determined during installation and should not be changed<br />

unless the Bright Field file is moved.<br />

The text to the left of the equal sign should always indicate the location of the Bright Field file. (Refer to<br />

the appendix on the Glyco <strong>DOC</strong> system file structure.)<br />

4. Inspect the image display for scratches and other artifacts.<br />

Open the Bright Field file, which is stored as the file se1000.brt.<br />

View the entire image.<br />

• If scratches are seen in the bright field image, then a new Bright Field image must be acquired<br />

using a new clean, scratch free Bright Field Plate. Replacement Bright Field Plates can be ordered<br />

from <strong>Bio</strong>-<strong>Rad</strong>.<br />

• If blotches, smudges or film are seen, then try to clean the plate.<br />

33


Maintenance and Troubleshooting<br />

4.5 IMAGER TROUBLESHOOTING<br />

Try all appropriate actions in the chart below for the imaging problem you are experiencing, before calling a<br />

<strong>Bio</strong>-<strong>Rad</strong> technical representative (Appendix D).<br />

Table 3<br />

Imager Troubleshooting<br />

Indication Problem Action<br />

Image is solid black in normal Imager is turned off. Turn on imager.<br />

palette and the acquire time is<br />

approximately 4 seconds. UV light has burned out. Replace UV light.<br />

Image is solid black in normal Loose or disconnected cable. Check for secure cable<br />

palette and the acquire time is<br />

connections at the computer<br />

0 seconds. and the imager.<br />

Image is mostly black in normal Filter Switch is between Check that the Filter Switch<br />

palette and the acquire time is positions. is at either 10 o’clock or 2<br />

approximately 20 seconds.<br />

o’clock. The switch should<br />

Neither Filter light is on,<br />

easily stop at these two<br />

but the power light is.<br />

positions. Forcing the switch<br />

should never be necessary.<br />

Image is mostly black in normal Gel was left in imager Remove gel when prompted<br />

palette, except for saturated areas. during background acquire. by the software during acquire.<br />

Do not select OK in the dialog<br />

box until the gel has been<br />

removed and the compartment<br />

closed.<br />

Acquired image is unusually Imager and/or camera board is Turn on Imager and PC a<br />

bright and of poor quality. not warmed up. minimum of two minutes<br />

prior to operation.<br />

Acquired image appears to Acquire time is too long. Reduce the acquire time.<br />

have a “snowy” background.<br />

Acquired image has bright Gel glass has lint or acrylamide Clean gel glass to remove all<br />

blob-like spots. particles. lint or acrylamide.<br />

34


Maintenance and Troubleshooting<br />

Table 3 (continued)<br />

Imager Troubleshooting<br />

Indication Problem Action<br />

Too little or too much of the Incorrect Image Rectangle. Reselect the Image Rectangle<br />

gel is being acquired.<br />

using the Select Image<br />

Rectangle command in the<br />

Image menu’s Acquire<br />

Options.<br />

Unable to acquire one side Mirror is out of position. Call for service.<br />

of the gel. When selecting the<br />

Image Rectangle, that side of<br />

the gel is also not visible.<br />

Acquired image is unusually Camera is out of focus. Call for service.<br />

blurry.<br />

4.6 SOFTWARE TROUBLESHOOTING<br />

4.6.1Memory Problems<br />

Glyco Doc analytical software works with large image files which take up a lot of hard disk and system memory.<br />

Its operations are impaired if there is less than 4MB of system memory available, or if the hard disk is<br />

full. Use About in the Help menu to check for available memory and disk space.<br />

MS Windows’ Virtual Memory -Permanent Swap File is a large hidden file which acts as a buffer for data<br />

storage. It serves the same function as RAM memory. We recommend you have a permanent swap file.<br />

Generally, it needs to be set-up only once. Closely follow the instructions in the MS Windows User’s Guide<br />

to determine if your system has a permanent swap file and, if not, how to create one.<br />

4.6.2Windows<br />

If a particular window can not be located on the screen, check to see if it is toggled off in the Glyco Doc<br />

Windows menu. A window will only be seen if it is toggled on in the menu. A checkmark, next to the<br />

option’s name, indicates the window is toggled on. A Band Table window, however, is only displayed if<br />

bands have been found.<br />

A window may be obscured by the Image Display . For example, if the Zoom Window is placed over the<br />

Image Display, it will be quickly covered by the display. Decrease the size of the image display until the<br />

Zoom Window is found. Then, move the Zoom Window away from the Image Display region by grabbing<br />

and dragging the title bar with the mouse. Because of its size, the Image Display may obscure any other<br />

windows in this way.<br />

35


Initialization File<br />

APPENDIX A.<br />

INITIALIZATION FILE<br />

The <strong>GLYCO</strong><strong>DOC</strong>.INI initialization file defines the default options an application automatically selects when it<br />

is first envoked. Because it is a text file, it can be easily edited by the user. The Glyco Doc Imaging initialization<br />

file specifies:<br />

• Window display defaults<br />

• Standard Lane Options<br />

• Colors<br />

• Band descriptions<br />

• Last Exposure Time<br />

• Bright File locations<br />

• Default data locations<br />

Note: The software must be restarted in order for the .INI file changes to take affect.<br />

A.1 WINDOW DISPLAY DEFAULTS<br />

The default settings shown below determine whether various windows appear at startup. Unless otherwise<br />

specified, “0” designates “off” and “1” designates “on” at startup.<br />

Image Histogram IGROn=1<br />

Densitometer<br />

Pixel Table<br />

Mouse Table<br />

Band Table<br />

Zoom Table<br />

DenOn=1<br />

DenMode=0 ; 0 for Marked region, 1 for Selected Lane<br />

TBVBlockOn=0<br />

TBVMouseOn=1<br />

TBVBandsOn=1<br />

TBVBandsMode=0 ; cycles through the various modes 0-8<br />

ZoomOn=1<br />

A.2 STANDARD LANE OPTION DEFAULTS<br />

Option default are the startup settings for the items in the Options menu:<br />

Reverse Palette ReversePalette=1<br />

Show Lane Center LaneStyle=0<br />

Show Saturated Pixels HiliteSaturatedPix=1<br />

Autoscale on image update AutoScale=1<br />

Show Associations ShowRows=1<br />

36


Initialization File<br />

A.3 COLORS<br />

Colors are designated by their Red, Green, and Blue contents. Some common colors are:<br />

• Red 255 0 0<br />

• Green 0 255 0<br />

• Blue 0 0 255<br />

• Yellow 255 255 0<br />

• Tourquise 0 255 255<br />

• Aqua 0 128 128<br />

• Blueberry 0 0 128<br />

• Magenta 255 0 255<br />

• White 255 255 255<br />

• Black 0 0 0<br />

• SaturatedPixsColor=255 255 0<br />

• IGRBkColor=0 255 255<br />

• IGRBorderColor=192 192 192<br />

• IGRRangeMarkColor=255 0 0<br />

• ZoomBorderColor=0 128 128<br />

• DenBkColor=0 255 255<br />

• DenBorderColor=0 0 255<br />

• MenuHelpBkColor=0 0 128<br />

• MenuHelpTextColor=255 255 255<br />

• BandNormalColor=255 0 0<br />

• BandSelectColor=0 255 0<br />

• BandHiLiteColor=0 255 255<br />

• BandHiSelColor=0 255 0<br />

• LaneNormalColor=255 0 255<br />

• LaneSelectColor=0 255 0<br />

• LaneHiLiteColor=0 255 255<br />

• LaneHiSelColor=0 255 0<br />

• LtGrayBackground=1<br />

• DenSolidBlack=0<br />

37


Initialization File<br />

A.4 BAND DESCRIPTIONS<br />

The value set in the Band Associations Dialog Box is the band association pixel distance. It determines<br />

whether or not two bands are associated.<br />

• BandLinkOverlap=6<br />

Band description settings are used by the Band Descriptions pulldown item.<br />

• G2=<br />

• G3=<br />

• G4=<br />

• G5=<br />

• G6=<br />

• G7=<br />

• G8=<br />

• G9=<br />

• G10=<br />

• G11=<br />

• G12=<br />

• GalNAc=<br />

• Glucose=<br />

• Galactose=<br />

• Fucose=<br />

• Mannose=<br />

• Sialic Acid=<br />

• GlcNAc=<br />

KeyBand specifies which name(s) will appear in the Standard Lane Analysis Dialog Boxes. It is the only<br />

descriptor needed for Standard Lane Analysis. Dp is the Degree of Polymerization for that band. The RF<br />

descriptors specify which bands to use for setting Rf values. Quantity is the pmol value to assign to the<br />

band named in the QuantBand description.<br />

38


Initialization File<br />

[Monosaccharides]<br />

• KeyBand=GlcNAc<br />

• RF0Band=GlcNAc<br />

• RF1Band=GalNAc<br />

• BandNames=GlcNAc 50.00,Galactose 50.00,Glucose 50.00,Fucose 50.00,Mannose 50.00,GalNAc<br />

50.00<br />

[Oligosaccharides]<br />

• KeyBand=G4<br />

• RF0Band=G12<br />

• RF1Band=G4<br />

• DP=4<br />

• Quantity=50.0<br />

• QuantBand=G4<br />

• BandNames=G2,G3,G4,G5,G6,G7,G8,G9,G10,G11,G12,G13,G14<br />

[Lane Descriptions]<br />

• Monosaccharides=<br />

• Oligosaccharides=<br />

• StandardsNames=Monosaccharides,Oligosaccharides<br />

A.5 LAST EXPOSURE TIME<br />

This reflects the last user-entered exposure time.<br />

A.6 BRIGHT FILE LOCATIONS<br />

The location of the Bright Field file (SE1000.brt) is given in the .INI file and therefore, should be placed in<br />

the directory specified in that file. The .INI specification for default Bright Field file is used for the bright field.<br />

A.7 DEFAULT DATA LOCATIONS<br />

File defaults, where files are saved to and important files located are:<br />

• DefaultDataDir=c:\face\data\<br />

• DefaultBrightFileDir=c:\face\data\<br />

39


Imager Specifications<br />

APPENDIX B.<br />

IMAGER SPECIFICATIONS<br />

Imaging Type:<br />

Detector Type:<br />

Light Source:<br />

Fluorescence, Transmittance<br />

CCD array, 753 x 244 pixels<br />

UV lamp filtered for maximum output at 368 nm<br />

Imaging Resolution: Vertical resolution of 320 µm<br />

Excitation:<br />

Imaging Area:<br />

Imager Dimensions:<br />

Interface:<br />

File Formats:<br />

Reports Formats:<br />

Minimum Computer &<br />

Display Requirements<br />

Wavelength maximum output at 368 nm<br />

10 x 10 cm cassette holder, 8 x 8 cm imaging area<br />

12.25 (H) x 7 (W) x 12 (D) inches<br />

ISA PC card (included)<br />

Exports TIFF, WMF, BMP, PCX, GIF, and PICT file formats<br />

Image, band data, densitometer, and integrated reports<br />

Computer: IBM ® -compatible 386 25 MHz computer; 256 gray scale or<br />

256 color video output<br />

Display: 800 x 600 pixel resolution<br />

Power requirements<br />

(switch selectable)<br />

120 volt setting: 100-120 V, 50/60 Hz; Fuse*: 250 mA, Time Delay<br />

240 volt setting: 220-240 V, 50/60 Hz; Fuse*: 160 mA, Time Delay<br />

*Note: User must ensure correct fuse is in place for selected voltage setting.<br />

Environmental<br />

Operating Temperature:<br />

Humidity:<br />

4° to 40° C<br />

0 to 95%, in the absence of condensation<br />

40


Warranty and Ordering Information<br />

APPENDIX C.<br />

WARRANTY AND ORDERING INFORMATION<br />

The Glyco Doc Imaging System is warranted for 1 year against defects in materials and workmanship. If<br />

any defects should occur during this warranty period, <strong>Bio</strong>-<strong>Rad</strong> Laboratories will replace the defective parts<br />

without charge. However, the following defects are specifically excluded:<br />

1. Defects caused by improper operation.<br />

2. Repair or modification done by anyone other than <strong>Bio</strong>-<strong>Rad</strong> Laboratories or their authorized agent.<br />

3. Use with other spare parts not specified by <strong>Bio</strong>-<strong>Rad</strong> Laboratories.<br />

4. Damage caused by deliberate or accidental misuse.<br />

5. Damage caused by disaster.<br />

6. Damage due to use of improper solvent or sample.<br />

For inquiry or request for repair service, contact your local <strong>Bio</strong>-<strong>Rad</strong> office.<br />

<strong>Bio</strong>-<strong>Rad</strong> Laboratories Belgium Italy Spain<br />

2000 Alfred Nobel Drive <strong>Bio</strong>-<strong>Rad</strong> Laboratories S.A.-N.V. <strong>Bio</strong>-<strong>Rad</strong> Laboratories S.r.l <strong>Bio</strong>-<strong>Rad</strong> Laboratories, S.A.<br />

Hercules, California 94547 Begoniastraat 5 Via Cellini, 18A Avda, Valdelaparra, 3<br />

Phone: (510) 741-1000 B-9810 Nazareth Eke 20090 Segrate - Milano Poligono Industrial de Alcobendas<br />

1-(800) 4-BIORAD Phone: 091-85-55-11 Phone: 02/21609.1 E-28100 Alcobendas (Madrid)<br />

1-(800) 424-6723 Fax: 091-85-65-54 Fax: 02/21609.399 Phone: (91) 661 70 85<br />

Fax: (510) 741-1060 or (900) 100 204<br />

1-(800) 879-2289 Fax: (91) 661-96-98<br />

Telex: 335-358<br />

Eastern Regional Office Peoples Republic of China Japan Switzerland<br />

85A Marcus Drive <strong>Bio</strong>-<strong>Rad</strong> Pacific (Beijing Office) Nippon <strong>Bio</strong>-<strong>Rad</strong> Laboratories KK <strong>Bio</strong>-<strong>Rad</strong> Laboratories, A.G.<br />

Melville, New York 11747 Yanshan Hotel Office Tower #1307 Sumitomo Seimei Kachidoki Bldg. Kanalstrasse 17<br />

Phone: (516) 756-2575 138A Haidian Road 5-3-6 Kachidoki 8152 Glattbrugg<br />

1-(800) 4-BIORAD Beijing 100086 Chuo-Ku Tokyo 104 Phone: 01-810-16 77<br />

1-(800) 424-6723 Phone: 2563146 Phone: 03-3534-7515 Fax: 01-810-19 33<br />

Fax: (516) 756-2594 or Fax: 2564308 Fax: 03-3534-8027<br />

1-(800) 756-4246<br />

Canada France Netherlands United Kingdom<br />

<strong>Bio</strong>-<strong>Rad</strong> Laboratories Ltd. <strong>Bio</strong>-<strong>Rad</strong> S.A. <strong>Bio</strong>-<strong>Rad</strong> Laboratories B.V. <strong>Bio</strong>-<strong>Rad</strong> Laboratories, Ltd.<br />

5149 Bradco Boulevard 94/96 rue Victor Hugo Fokkerstraat 10 <strong>Bio</strong>-<strong>Rad</strong> House<br />

Mississauga, Ontario L4W 2A6 B.P. 220 3905 KV Veenendaal Maylands Avenue<br />

Phone: (416) 624-0713 94203 Ivry Sur Seine Cedex Phone: 08385-40666 Hemel Hempstead<br />

1-(800) 268-0213 Paris Fax: 08385-42216 Hertfordshire HP2 7TD<br />

Fax: (416) 624-3019 Phone: 01-4960-6834 Phone: 0442-232552<br />

Fax: 01-4671-2467 0800-181134<br />

Fax: 0442-259118<br />

Australia Germany New Zealand<br />

<strong>Bio</strong>-<strong>Rad</strong> Laboratories Pty., Ltd. <strong>Bio</strong>-<strong>Rad</strong> Laboratories GmbH <strong>Bio</strong>-<strong>Rad</strong> Laboratories Pty., Ltd.<br />

Unit 11 Heidemannstrasse 164 Unit 15 Poland Court<br />

112-118 Talavera Rd. Postfach 45 01 33 21 Poland Road<br />

P.O. Box 371 D-8000 Munchen 45 P.O. Box 100-051<br />

North Ryde Phone: 089 318 84-0 North Shore Mail Centre<br />

New South Wales 2113 Fax: 089 318 84-100 Glenfield, Auckland 10<br />

Phone: 02-805-5000 Phone: 09-443 3099<br />

008-224-354 0800-805 500<br />

Fax: 02-805-1920 Fax: 09-443 3097<br />

Telex: 79070166<br />

Austria Hong Kong Scandinavia<br />

<strong>Bio</strong>-<strong>Rad</strong> Laboratories Ges.m.b.H. <strong>Bio</strong>-<strong>Rad</strong> Pacific Ltd. <strong>Bio</strong>-<strong>Rad</strong> Laboratories<br />

Auhofstrasse 78D Unit 1111, 11/F., New Kowloon Plaza Kanalvagen 10C<br />

A-1130 Wien 38 Tai Kok Tsui Road 19461 Upplands Vasby<br />

Phone: 0222-877 89 01 Tai Kok Tsui , Kowloon Phone: 46 (0)8 590-734 89<br />

Fax: 0222-876-56-29 Phone: 7893300 Fax: 46 (0)8 590-717 81<br />

Telex: 13-6565 Fax: 7891257<br />

Telex: 42814<br />

41


Warranty and Ordering Information<br />

WARRANTY INFORMATION<br />

Model: ______________________________________________________________________<br />

Serial Number:________________________________________________________________<br />

Date of Delivery: ______________________________________________________________<br />

Warranty Period: ______________________________________________________________<br />

ORDERING INFORMATION<br />

Prices and part numbers are subject to change. Call for current prices.<br />

Catalog<br />

Number<br />

Description<br />

170-6555 Glyco Doc Imager, 100/240 V, includes Glyco Doc imager, Glyco Doc analytical<br />

software, and Glyco Doc imager interface card and cable.<br />

170-6557 Glyco Doc Imaging System, 100/240 V, includes Glyco Doc imager, Dell 486<br />

computer with 8 MB RAM, 1.08 GB internal hard drive, and 230 MB internal optical<br />

drive, pre-loaded Glyco Doc analytical software, DOS 6.0, MS Windows 3.1, preinstalled<br />

Glyco Doc imager interface card, 15: Enhanced Super VGA monitor,<br />

keyboard, MS mouse, and HP Laserjet printer.<br />

170-6559 Glyco Doc Analytical Software<br />

170-6575 UV Lamp (1)<br />

170-6642 Test Plate (1)<br />

170-6641 Bright Field Plate (1)<br />

170-6576 Glyco Doc Glass Back Plate<br />

42


?O2@6K ?O2<br />

?O2@@@@@@@@@6K<br />

?O2@@@@<br />

?O2@@@@@@0M?I4@@@@@@6K<br />

?O2@@@@@@0M<br />

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<strong>Bio</strong>-<strong>Rad</strong><br />

Laboratories, Inc.<br />

Life Science<br />

Group<br />

2000 Alfred Nobel Drive<br />

Hercules, California 94547<br />

Phone: (510) 741-1000<br />

Fax: (510) 741-1060<br />

<strong>Bio</strong>-<strong>Rad</strong> Laboratories Main Office, 2000 Alfred Nobel Drive, Hercules, California 94547, Ph. (510) 741-1000, Fx. (510) 741-5800<br />

Also in: Reagents Park, Australia, Ph. 02-9914-2800, Fx. 02-9914-2888 Wien, Austria, Ph. (1) 877 89 01, Fx. (1) 876 56 29 Nazareth, Belgium, Ph. 09-385 55 11, Fx. 09-385 65 54<br />

Mississauga, Canada, Ph. (905) 712-2771, Fx. (905) 712-2990 Beijing, China, Ph. (01) 2046622, Fx. (01) 2051876 Copenhagen, Denmark, Ph. 39 17 9947, Fx. 39 27 1698<br />

Espoo, Finland, Ph. 90 804 2200, Fx. 90 804 1100 Ivry sur Seine Cedex, France, Ph. (1) 49 60 68 34, Fx. (1) 46 71 24 67 München, Germany, Ph. 089 31884-0, Fx. 089 31884-100<br />

New Delhi, India, Ph. 91-11-461-0103, Fx. 91-11-461-0765 Milano, Italy, Ph. 02-21609.1, Fx. 02-21609.399 Tokyo, Japan, Ph. 03-5811-6270 Fx. 03-5811-6272 Veenendaal,<br />

The Netherlands, Ph. 0318-540666, Fx. 0318-542216 Auckland, New Zealand, Ph. 09-443 3099, Fx. 09-443 3097 Kowloon, Hong Kong, Ph. 7893300, Fx. 7891257 Singapore,<br />

Ph. (65) 272-9877, Fx. (65) 273-4835 Solna, Sweden, Ph. 46 (0) 8 735 83 00, Fx 46 (0) 735 54 60 Madrid, Spain, Ph. (91) 661 70 85, Fx. (91) 661 96 98 Glattbrugg, Switzerland,<br />

Ph. 01/809 55 55, Fx. 01/809 55 00 Hemel Hempstead, United Kingdom, Ph. 0800 181134, Fx. 01442 259118<br />

SIG 020996<br />

Printed in USA<br />

400 6059 Rev C

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