I have been staining murine lung tissue with Immunofluorescence to collect B cells using Laser Capture Microdissection. The staining looks great, but when I would test the IR spots to collect cells, the laser would fire an enormous spot for the tissue (around 40 um). I played around with the power, and duration of the laser but nothing seemed to decrease the spot size. If it did, the laser would not be powerful enough to actually cut through the tissue and lift my cells.
So I decided to try and collect some cells anyway with what I could do. The spot size was huge, and seemingly destroyed not only the tissue around it but the cells I wanted too. When i brought the cap to QC the flourescence was gone, so I'm assuming the cells got fried by the laser.
Does anyone know how to troubleshoot this? I am using regular glass slides cut from frozen OCT sections, and the macro arcturus caps. I ordered some of the HS caps, so we will see if those make a difference. We are also using the arcturus XT microscope and software.
The MMI CellCut laser microdissection uses an UV laser with very low power but high pulse rate to create a very thin cutting line with sub-micron precision not damaging the cells of interest or the surrounding tissue.
Please check the website (inlcuding a short video) on how this works:
https://www.molecular-machines.com/products/cellcut
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