I recently stained some cells with CFSE and used an adoptive transfer method. I'm now looking for those cells and any changes in tissue morphology, and opted for a standard H&E stain from the histology service (I did not perform it myself since I do not have the equipment for sectioning). My tissue is highly autofluorescent in the FITC range! The tissues were formalin-fixed and paraffin-embedded (the paraffin gives some fluorescence background, but not much). I have unstained sections remaining, but need a good, NON-fluorescent way to bring out tissue morphology. Any suggestions?
The observation of HE stain, you can just use optical microscope. You do not have to do excitation of FITC....
If I were only interested in the H&E, I could use optical only. However, I need to see BOTH the FITC/CFSE fluorescence as well as the tissue morphology. Since H&E gives me high background fluorescene, what other options should I consider?
Now I understand what you mean. You mean HE stain affects to FICT and avoid to do that, right? I normally use the phase contrast microscope for observation of whole tissue without staining in that case. Or instead of using FITC, other fluorescences....
Phase contrast would be a second choice (and I'm not sure the microscope has the right lenses). We are using CFSE (which emits in the same basic range as FITC) to label some cells that we want to see later in tissue, and determine whether a treatment physically affected the cell survival or tissue morphology. The problem is that eosin is also fluorescent in the FITC range. I was thinking about maybe Masson's Trichrome but I have no experience with it personally-has anyone observed it to be fluorescent?
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