I seem to be having a problem with imaging at high magnification. I need to image at 60X to evaluate spine densities but even at 40X, the objective seems to need to be so close to the coverslip that there is pressure which causes the sample to float away in the mounting media. Has anyone else had this issue? I would appreciate any suggestions/advice. I am only trying to image through a 200um thick section so this should not be a problem at all yet here we are.
We had a barrier of bluetac (putty) on a glass slide with a tiny opening to insert a pipette, then placed the specimen in the middle (have a drop of RapiClear on the glass slide and the top of your section when you place it to avoid air bubbles) and then we pressed the willco dish on top evenly so that it touched the bluetac and the sample and held it in place. Then we filled the newly created "chamber" with RapiClear and sealed the opening with KwikSil.
Does that help?