could you please help me with the following questions?
1. How to capture single cells or tissue pieces of only 40 microns diameter (Arcturus microscope, PEN membrane glass slides, frozen sections, stained with Ambion kit):
1.1 Which settings for the IR-laser (diameter, power, duration)?
1.2 Do I have to use the UV-laser, if yes with which settings (tab length, UV cut length)?
2. I have problems with repeatability: e.g. first I capture material, but then, only a few minutes later, I don't get any material off the slide.
3. Does it affect the downstream analysis (RT-PCR) if the material gets grey (like burned) by the laser? That is often the case if I use settings like IR: 30 microns spot diameter, 70 mw power, 28 msec duration; UV: 10 microns tab lenght, 500 microns UV cut length).
4. Is it normal that the piece on the cap has not got the same size and shape as the "hole" in the remaining material on the slide?
Thank you in advance!