1. Cyclone System Overview How it works Applications Features & Benefits

2. What does the Cyclone  do? Cyclone makes images of flat radioactive samples: 3 H, 14 C, 32 P, 35 S 125 I, 18 F, etc. 3 H, 14 C, 32 P, 35 S 125 I, 18 F, etc. Gels, blots, tissue sections, gene arraysGels, blots, tissue sections, gene arrays Film is the traditional means of imaging

3. How does Cyclone work? BaFBr:Eu2+ BaFBr:Eu3+ BaFBr:Eu2+*

4. Cyclone –Parts and Accessories Cyclone Storage Phosphor Scanner Computer System 2 Storage Phosphor Screen(medium size MP screens) Scanning Carousel(for medium size screens) SCSI II Interface Card SCSI II Interface cable OptiQuant software Operation Manuals

5. Cyclone -Specification Physical Specifications Environmental Requirements Computer Requirements Performance Specifications

6. Physical Specifications ScannerDimensions Height11inches(27.9cm) Width17inches(43.2cm) Depth10inches(25.4cm) Weight37lbs(17kg) Screens Small5x7.6inches(12.5x19.2cm) Medium5x10inches(12.5x25.2cm) Large5x17inches(12.5x43.0cm)

7. Environmental Requirements Electrical Voltage 100 to 240 VAC at 50 or 60 Hz Overvoltage Category II Pollution Degree 2 ClassI Current 0.9A at 100-130VAC 0.6A at 200-240VAC Fuse Rating(5x20mm fuses) 1A SB-125V at 100-130VAC 0.63A T-250V at 200-240VAC Power 100 Watts Temperature 60 to 90 F (15-32C) Humidity 5 to 95% relative humidity, non- condensing (for indoor use only) Altitude To be used under 6500ft(2000m) consult factory for higher altitude

8. Computer Requirements(minimum) CPU Pentium 75MHz RAM32MB Hard Disk 1 GB VideoSVGA(1024x768recommended) Scanner Interface SCSI II Operating System Microsoft Windows 95 Image File Size (full area of large screen) 61.5MB at 600dpi 15.5MB at 300dpi 7MB at 200dpi 7MB at 200dpi 4MB at 150dpi 4MB at 150dpi

9. Performance Specifications Sensitivity (1hr exposure) <2DPM/mm2 for C14 / <2DPM/mm2 for S35 <2DPM/mm2 for C14 / <2DPM/mm2 for S35 <1DPM/mm2 for P32 / <1DPM/mm2 for I125 Pixel Resolutions 600 dpi (42um pixel size) / 300 dpi (85um pixel size) 200 dpi (127um pixel size) / 150 dpi (170um pixel size) Spacial Resolution (2.5 line pairs/mm) 0.33 CTF for C14 on SR screen 0.70 CTF for H3 on TR screen Uniformity +/-5% over scan area Linear Dynamic Rang 5 orders of magnitude (1-100,000) Linearity +/-5% standard deviation for dynamic range Scan Time (full area of large screen) -9 min at 600dpi / -5 min at 300dpi -4 min at 200dpi / -3 min at 150dpi Screen Erasure Time 0.5 to 1 minute

10. Cyclone- Process of Storage Phosphor Imaging 1.Erase the storage phosphor screen just prior to exposure 2.Insert the sample and the screen into an exposure cassette 3.Expose the screen for an appropriate length of time 4.Remove the exposed screen from the cassette and clip it on to the Cyclone scanning carousel 5.Insert the carousel into the scanner and start the image acquisition from the OptiQuant software 6.Analyze and store the image using the OptiQuant software

11. Handling the Storage Phosphor Screens Light Sensitive-white light,UV,Bright fluorescent light Physical Damage –portion of the screen to be unusable Precautionary Handling Guidelines Do not bend,scratch,smudge or apply pressure to the storage phosphor screens. Keep phosphor screens away from hot or humid locations Keep phosphor screens away from direct contact with moisture Keep phosphor screens away from direct sunlight and ultraviolet rays Do not apply direct light to a phosphor screen that has been exposed but not yet scanned Dry the samples well and wrap them in protective film to prevent moisture damage and contamination to the screen Handle screens by the edges, the back or the taped ends

12. Cyclone-The Storage Phosphor Screen Size and Grade of the Storage Phosphor Screens Description of the Storage Phosphor Screens Handling the Storage Phosphor Screens Erasing the Storage Phosphor Screens Cleaning the Storage Phosphor Screens Storing the Storage Phosphor Screens Disposal the Storage Phosphor Screens

13. Choice of Phosphor Screen Sizes Small 5” X 7.6”, Medium 5” X 10”, Long 5” X 17”

14. MP, MultiPurpose and New MS, MultiSensitive for moisture resistance & durability SR, Super Resolution for best separation of bands ST, Super Sensitive for low activity samples Choice of Screen Grades TR, Tritium Sensitive for tritium labeled samples

15. Description of the Storage Phosphor Screens Phosphor(imaging) Side: White or Blue Protect from light, dirt, contamination and rough handling Back Side: Black Wet Sample/Acetic acid vapor/organic solvent vapor/Strongly acidic/alkaline solution

16. Erasing the Storage Phosphor Place the screen against the white light source with the phosphor side(imaging side) facing the light. 30-60s Actual Erasure Time: Energy of the original isotope Original image exposure time Whether the image was ever scanned Intensity of the white light source

17. Cleaning the Storage Phosphor Screens MP,SR, and ST types of phosphor screen can be cleaned to remove small amounts of radioactive contamination,dust,dirt,fingerprints and static electricity TR type of phosphor screen: Don’t attempt to clean Storing the Storage Phosphor Screens First the screen should be decontaminated and cleaned Store the screens at room temperature in a dry location away from all sources of light, beta,gamma and x-ray emission

18. Disposal of Storage Phosphor Screens

19. Exposing the Storage Phosphor Screen Preparing the Sample Preparing the Storage Phosphor Screen Defining the Exposure Loading the Exposure Cassette

20. Preparing the Sample Prepare the samples as you would for typical film autoradiography Dry gels for best resolution and for less self absorption Wrap all samples a 10um polyvinylidene chloride film when using the MP, SR, or ST screen types immediately prior to exposure To detect tritium, place the sample directly in contact with the phosphor side of TR screen Preparing the Storage Phosphor Screen Make sure that the phosphor screen is free of any existing image,dirt,fingerprints,damage or contamination

21. Defining the Exposure The optimum exposure time will depend on four factors The energy of the isotope in the sample (the higher the energy the shorter the exposure time required) The amount of radioactivity in the sample ( the greater the activity the shorter the exposure time required) The sensitivity of the phosphor screen using (for example, MP screen type may require a longer exposure time with 125I than an ST screen) The required quantitative integrity of the resulting image(the longer the exposure time, the better the picture quality due to the increased amount of data in the image)

22. Loading the Exposure Cassette Open the cassette and place the sample material in the cassette,wrapping it in plastic wrap Make sure that the sample is lying flat in the cassette and place the phosphor screen, imaging side down, on top of the sample Position the screen such that the sample image area are at least1/4 inch (6mm) away from the edges and taped ends of the screen. Mark on the tape near the upper left corner of screen With the sample and the phosphor screen positioned together in the exposure cassette, latch the back of cassette into the frame Leave the closed cassette at room temperature or in a refrigerator or –20C freezer for the amount of the time determined for an optimum exposure

23. Scanning the Storage Phosphor Screen Preparing the Scanner Loading the Storage Phosphor Screen Scanning the Exposed Storage Phosphor Screen

24. Preparing the Scanner Turn on the power to the Cyclone Scanner before apply power to the computer Make Sure Communicating with the OptiQuant software If you use the scanner frequently, you may choose to leave the power to the scanner on all of the time

25. Loading the Storage Phosphor Screen 1. Turn off the lights 2. Open the exposure cassette and remove the screen 3. Open one of the clamps on the scanning carousel to lift the gripper bar away from the body of the carousel drum 4. Carefully insert one taped end of the screen,under the open clamp on the carousel and close the clamp. Be sure that the phosphor side will face away from the carousel drum for scanning 5. Wrap the screen around the drum of the carousel, secure the free end under the second clamp 6. Open the lid of the Cyclone scanner and place the carousel into the scanner 7. Close the scanner lid

26. Scanning the Exposed Storage Phosphor Screen Define the image acquisition protocol and start the acquisition Open the top lid and remove the carousel from the scanner when the Scanning light stops blinking Erase the screen before using it again

27. Data Acquisition Cyclone Control Room Protocol Menu Status Menu Acquisition Menu

28. Maintenance Cleaning the Exposure Cassette Cleaning rhe Image Eraser Cleaning the Scanner Cleaning the Storage Phosphor Screens