BioChemi Image Acquisition and Analysis System Installation and User Instructions BioImaging Systems
BIOCHEMI IMAGE ACQUISITION AND ANALYSIS SYSTEM INSTALLATION AND ACQUISITION MANUAL Table of Contents Section 1: Introduction ...................................................................................... 3 Section 2: Safety Information............................................................................ 5 Section 3: BioChemi System Installation ..........................................................
Section 1 - Introduction Congratulations on your recent BioChemi purchase! You have selected the most comprehensive BioImaging System available on the market today. UVP strives to keep up with the advancements in technology, and wishes to assist you in any way possible. How to use this manual: The following compilation is intended to help the user install the BioChemi system in a step by step procedure. The first section will discuss setting up the computer hardware (computer boards, monitor, printer).
. EpiChemi II Darkroom • • Overhead 254 nm UV, 365 nm UV, and fluorescent white light • Enables the user to view the setup of the experiment with the white light on while the darkroom door is open. • Concealed viewer with UV blocking safety glass. • Enables the user to view the experiment while in progress, while protecting from UV exposure. Completely light tight darkroom. • • Retractable, sliding transilluminator tray. • • WWW.UVP.
Section 2 - Safety Information NOTE: All UVP High Performance Transilluminators are powerful sources of Ultraviolet (UV) radiation that can cause damage to unprotected eyes and skin. Be sure all personnel in the area are properly trained and protected before operating any unit that uses UV radiation. It is further recommended that the transilluminator be installed and operated in a darkroom where access and exposure is limited while the unit is in operation.
Section 3 - BioChemi Installation 1. EpiChemi II Darkroom Components of the EpiChemi II Darkroom are described here. Please refer to your packing slip for components included in shipment. A. Camera B. Camera mount C. Mounting knobs and washers D. Zoom lens E. Rubber light tight seal F. Overhead light switch G. 5-position filter control H. Transilluminator power switch I. Master darkroom power switch I. Darkroom Door J. Sample Viewer K. Fold Away Chemi Tray L. Compression Door Lock M.
1. Unpack the EpiChemi II Darkroom and Benchtop transilluminator from the packaging materials. 2. Assemble the two together by placing the transilluminator on the sliding tray of the darkroom. 3. Plug the Transilluminator in by using the internal power cord found inside the darkroom. *Note: The transilluminator is powered from the inside of the darkroom, which allows the transilluminator to be controlled from the darkroom switches. *NOTE: WHEN IMAGING, MAKE SURE THAT THE VIEWING WINDOW IS CLOSED.
2. BioChemi Cooled CCD Assembly and Installation 1. The camera AC power adapter should be plugged to the wall • with the power cable provided. Power Connection (plug supply into wall outlet) power 2. There are two BNC (video) connector plugs, Rx and Tx, that plug into the video sockets on the camera. Connect the Tx from the back of the camera to the Rx connection on the video board installed in your computer.
3. PCI Image Acquisition Board Installation Note: The Capture Board is relatively easy to install. If, however, you have reservations about installing it, we recommend that you ask an experienced technician to do so. If someone is not available, call UVP technical support for further instructions. 1. Be sure the power to your computer, monitor and other peripheral devices are turned off. You can cause damage to the components in your computer if you attempt to remove or insert boards with the power on. 2.
4. To remove the computer cover, remove the screws from the back of the case and slide the case off the computer. 5. Install the Capture Kit board into the PCI slot. Take the new board out of the antistatic bag. Hold it by the edges to prevent damage to the components or the edge connectors (the gold-colored “fingers” that extend from the board). 6. Still holding it by its edges, gently but firmly seat the “fingers” into the PCI slot.
4. Lenses The BioChemi system comes equipped with zoom and fixed lenses. Zoom lens features: Components of the zoom lens are as follows: 1. Top Ring: f-stop. This ring controls how much light the camera will see. 2. Middle Ring: Zoom. This ring controls the field of view that the camera will image. 3. Bottom Ring: Focus. This will adjust the sharpness of the image the camera will image. 4. Diopter: A +1 diopter is included. This is necessary for the lens to stay in focus as you zoom in and out.
5. Mounting the Camera 1. To attach the camera bracket to the top of the EpiChemi II darkroom, place the black rubber gasket over the four screw holes and align the camera mount. Screw in the (4) provided screws to attach the camera mounting bracket and light sealing gasket to the top of the darkroom. • Camera Mount • Rubber Gasket 2. Unpack the camera and zoom lens from their protective boxes and remove the protective plugs. 3. Screw the zoom lens into the end of the camera housing. 4.
5. Elevate 1/8” and screw the mounting bolts into the camera. Tighten, straighten, and secure. Make sure the zoom lens and the light sealing gasket are situated properly. WWW.UVP.COM 13 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
6. Installing LabWorks and Capture Driver Computer The computer power cable should plug directly into a power strip or surge protector. Find and locate the mouse, keyboard, and other accessories, plug in and secure all connections. Start up the computer once all of the connections have been made and the power is attached. NOTE: If the computer has LabWorks pre-installed, skip the installation procedures. Installation Procedure 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14.
17. 18. 19. 20. 21. 22. 23. 24. 25. Click Device Manager Tab Look for ‘Other Devices’ or ‘Sound Video Game Controllers’ category Double click them to open the lists Look for PCO PCI capture board 525/520 under Sound Video Game Controllers Or Look for Multimedia Audio/Video device under Other Devices Double click on it (if neither exists, go to #33 below) Choose Driver tab Click update driver Go up to #3 and continue installation ****************************** 26. 27. 28. 29. 30. 31. 32.
7. Printer and Security Dongle Unpack the Thermal Printer (if purchased) and place this alongside the computer. The printer is supplied with a separate instruction manual giving full operation and service details. The printer driver is preloaded on computer purchased with systems. If you need to install the driver onto a separate computer, please see the end of this section.
3. AC in Connector (power input) • Connect to a wall outlet using the AC power cord supplied The printer power cable should plug directly from the printer into the wall or a power strip. The data cable for the printer is a 25-pin parallel connector. This 25-pin connector will plug into the female equivalent in the back of the computer. dongle Find the open 25 pin female slot, this is the printer port. As soon as this port has been identified, locate the Security Dongle, supplied with the software package.
Section 4 - BioChemi Capture A. Non-Fluorescent Samples (Protein Gels) Transmitted White Light • Coomassie Blue • Sliver Stain • SYPRO Red • SYPRO Orange NO INTEGRATION NECESSARY Select the appropriate filter for the stain being used (See Appendix A for Filter Recommendations). 1. 2. Open up LabWorks. Click on the Camera Icon. 3. The Minimal Dialogue window will display. Stop/Capt When you click Stop/Capt, the Options button will become available. Click on the Options button. Options Button WWW.UVP.
4. Under the Integration tab, select No Integration (On-Chip Integration will be unavailable, Sequential Integration and Dynamic Integration should not be selected). Integration Tab WWW.UVP.COM 19 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
9. Check the Preview tab to make sure there is at least 55 Millisec entered. Numbers smaller than this will result in the inability to see a live preview as it is faster than the lower limit of the camera. Preview Tab Capture Button 10. Place the UVP target or printed text on the transilluminator for focusing. 11. To zoom in, turn the zoom lens all the way to the right and adjust the bottom for fine focus. 12.
B. Fluorescent Gels (DNA/RNA Gels/Some Protein) Transmitted UV Light • Ethidium Bromide • SYBR Green • SYBR Gold • Fluorescein • Rhodamine • Texas Red • Sypro Orange • Sypro Red Usually requires integration to make stained bands appear. To increase the brightness of the bands, increase the integration time. Select the appropriate filter for the stain being used (See Appendix A for Filter Recommendations). 1. 2. Open up LabWorks. Click on the Camera Icon. 3. The Minimal Dialog window will display.
4. From the Preview tab, click on the Start Preview button. Preview Tab Start Preview button WWW.UVP.COM 22 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
5. Click the Integration tab and make sure that No Integration, Sequential Integration and Dynamic Integration are unchecked. Integration Tab 5. Go to the Acquire tab, and adjust the exposure time to the desired amount of time. Acquire Tab WWW.UVP.COM 23 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
6. If you want to preview and acquire for the same exposure, the Lock Exposure Times needs to be checked in the Acquire menu. 7. Place the UVP target or printed text on the transilluminator. 8. Zoom in all the way and focus on the lettering on the gel cutter. 9. Adjust the aperture on the zoom lens until you can see your bands without saturating the image. Oversaturated areas in the preview will turn red. Make sure there is no red in the image before capture.
C. Chemiluminescent samples (Films and Membranes/Plates) Emitted light in a light tight environment. • • The time to capture chemiluminescence with the BioChemi system will depend on several factors, but typically is equivalent to the time it would take for the same sample on film. Always make sure to start integration immediately after the substrate has been added, especially since some substrates can degrade quickly (we recommend BioWest Extended Duration substrate). 1. 2. 3. 8.
10. Click on the Options button and the following options will display. 11. Depending on your sample and your experience with it, you have three modes of integration: No Integration, Sequential Integration and Dynamic Integration WWW.UVP.COM 26 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
No Integration – straight accumulation of signal on the chip. The image captured is representative of all of the information on the chip at the end of the capture time period. Application: Acquiring blots with known exposure times. 1. 2. With the Options box open, click on the Start Preview button. If not already selected, click on the Acquire tab (On Chip, Sequential Integration and Dynamic Integration should not be selected) and adjust the exposure time to the desired amount of time.
Sequential Integration – Sequence of images captured from multiple onchip integrations of increasing duration. Can be played back as a Digital Video Playback movie. Pre-set menu or programmable times available. Application: Low signal, limited duration substrates with high background (ECL). 1. Click the Integration tab. 2. Select Sequential Integration. Integration Tab Sequential Integration 3. Click Sequential Settings button.
Regular Intervals– Enter total amount of time and total number of frames you want to capture. This will produce X frames of T/X seconds each (T = Total Time, X = # of frames), and will play back in sequence. Regular Intervals Click OK after intervals are selected. This will return you to the Integration tab. Click Capture to capture the image. WWW.UVP.COM 29 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
Irregular Intervals – Select from the menu or create your own by entering in irregular intervals. Resulting frames will play back in sequence in “real time”. Irregular Intervals To define Irregular Intervals: 1. Create a new name in the Name field. 2. Click the Add button. 3. Enter exposure times in the Minutes, Seconds, or Milliseconds fields. 4. Click the Update button. 5. Click OK to close the Sequence Integration window and return to the Integration tab. 6. Click Capture. WWW.UVP.
Dynamic Integration Auto Exposure – Automatic accumulated exposure of multiple on-chip integrations before image saturation. Application: Immunoblot protocols using enhanced luminol substrates with high sensitivity and extended duration (BioWest, SuperSignal) 1. With the Integration tab open, check the Dynamic Integration, Auto Exposure, and Saturation Warning boxes. 2. Enter the amount of time you want to expose each frame. Each frame will be added to the one before it in an arithmetic progression. 3.
Dynamic Integration Total Time Exposure – Accumulated exposure of pre-set, on-chip integrations over a specified time interval with automatic Digital Video Playback (DVP) of stacked images. Application: Blotting with substrates of high signal but limited duration and low background (ECL Plus). 1. WWW.UVP.COM With the Integration tab open, check Dynamic Integration, Auto Exposure, and Total Time Exposure. 2. Click on the Total Time Settings button and the Dynamic Integration window will open. 3.
4. 5. WWW.UVP.COM stacked when played back through the Digital Video Playback mode. Click Capture. This automatically captures any live image before the brightest pixel reaches saturation. For longer exposures, it is best to also check the Integration box and set the seconds for approximately 1/10 of the total expected exposure time (i.e. 10 minutes = 60 seconds). This allows the CCD chip to be read every interval selected, displaying the image as it exposes over the entire integration time.
Section 5: Technical Support Before you contact UVP’s Technical Support for further assistance, please be prepared to provide the following information: Your registration information: product serial number (*), your name, phone number, organization name, mailing address, and e-mail address. The software version number (*) and system information as applicable. A description of the problem, and any relevant information regarding prior technical support assistance.
Appendix A – Recommended Stains Filter Emission Ethidium Bromide Ethidium Bromide Coomassie Blue SYPRO Orange Coomassie Blue Coomassie Blue SYBR Gold SYBR Gold SYPRO Red Texas Red Rhodamine SYBR Green GFP SYBR Green Fluorescein WWW.UVP.COM 35 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
Appendix B – Troubleshooting Guide Cannot capture Image. When selecting the Camera icon on the toolbar, there is a tab named Signal tab, which has an option called External Trigger. Make sure this option is NOT checked. The only way to capture an image if this option is checked is to press any key on the keyboard, this acts as the external trigger. Cannot view live preview, or image shows red or gray.
Appendix C – Maintenance Changing Bulbs in the EpiChemi II: Remove the Transilluminator to prevent any scratching to the glass surface. Tilt the darkroom back and use a 5/16-inch socket screwdriver or nutdriver to remove the 4 nuts holding the bulb cover in place. The 4 nuts to be removed: Removed light filter cover. WWW.UVP.COM 37 US Technical Support: 1.800.452.6788 UK Technical Support: +44.0.1223.
Remove bulbs by rotating the bulbs, approximately a ¼ turn, and pulling at the same time, much like replacing fluorescent overhead bulbs. Then replace the bulbs in the same configuration that they were originally installed; UV bulb, White bulb, and UV bulb configuration. Replace the cover and test lighting with the outside switch on the EPICHEMI II. *Note: When testing bulbs, be sure to wear proper UV protecting eyewear and any other appropriate personal protective equipment.
