FLUROIMAGER 595

This page will give general information on how to turn the scanner on, do a basic scan, and visualize the image. For more advanced applications, look at other protocols or read the manuals located next to the scanner.

Gel Scan:

under the start icon go to programs click on flora on ("scanner control" opens)
- To turn the scanner off, close the "scanner control" program
click on door open
put glass gel tray grid with gel into machine
on the computer screen highlight grid compatible to gel area
click setup
set the PMT voltage (500-900)
- A higher number is more sensitive, but may show more background
set the pixel size
- 100 micron gives higher resolution, but 4X the file size of 200 micron. We normally use 200 micron.
Leave the resolution at 16 bit.
Detection sensitivity
- High sensitivity gives a superior scan, but can take 4X as long
- If you are doing quantitation or AFLP you must use high, otherwise you can use normal.
set calibration 530df30
set orientation (click on appropriate)
click ok
click scan
Name the file and put it in the appropriate directory and click Save.

Adjust the Display:

(Directions are in general, there are many ways to adjust the display)

click on Grey/Color Adjust (Blue highlighted button to the right) (collects pixel values from 0-10,700 relative fluorescence units (rfu))

high (black) and low (white) display levels of images can be changed independently or together. While the color is being adjusted, everything that is green on the image is black, and everything purple is white.

high: pointer at top of bar (higlighted blue at left), hold left mouse button down, when image begins to turn green, stop, move pointer back up until all the green places are black. Press "Set Range".

low: pointer at bottom of bar, move mouse up the bar until background of image is purple but spots are not. Press "Set Range"

To reset the adjustment back to the original scan type in 999999 in the "Hi" box and 0 in the "Lo" box.