Dear all,
I am finding that some of the spleen sections that I make at the cryostat and attach to my glass slides, regardless of the tissue NOT demonstrating any cuts during the slicing, show many zebra like \ segmented cuts in the organ section after a DAPI 5 minute incubation.
Under a conventional microscope where I can see the DAPI, the spleen sections demonstrate these holes \ cuts inside the spleen section which are all over the organ. This is a pattern that by eye I cannot see when cutting the sample at the cryostat and find it so frustrating because I am not sure what I am doing wrong. This has occurred to cryomolds that previously did not show this pattern but eventually do, and I need to continue cutting to find the "not ruined" samples. However, it could be some detail that I am missing.
I would be forever grateful for your advice, as the self-doubt is driving me crazy!
Rebecca
This is maybe due to microchatter. Vibration of the blade because of too fast cutting, too low temperature, loose knife or tissueholder, too rough trimming, dull knife. Check if all parts are fixed, cut more slowely, cut more thin slices after trimming, take a fresh knife.
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