5.2.7.9. Close the iris under the Common tab. Go to reflected light under microscope tab. There are several buttons under fluorescence with filter names. Laser refers to the cutting laser, and cutting is only performed in brightfield.
5.2.7.10. Click on a filter name to open the reflected light iris and use filter light.
5.2.7.11. Hit measure to automatically adjust exposure time. Do not over expose your sample as this will decrease fluorescence signal. Limit the time that the sample is exposed to light. Analog gain is 3-4 dB, exposure depends on filter selection.
5.2.7.12. Click on advanced settings. Live Image window opens showing a histogram. O-255 refers to intensity, RGB are binned.
5.2.7.13. For saturation in histogram beyond 255, adjust intensity using by XCite wheel, exposure time, reducing gain.
5.2.7.14. To optimize histogram, drag the line by grabbing at the top and placing where light peak ends or bottom and placing were light path of overlapping light ends. Increase exposure time until optimized. Save settings under live image.
5.2.7.15. Changing cameras to black and white may increase sensitivity. Histograms are optimized the same way.
5.2.7.16. Adjusting the gain: XCite wheel, reduce gain under microscope tab, or reduce exposure time.