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| Hi! I am trying to stain both adults and embryos on polylysine coated slides. I have been putting adults on coated slides and pressing down slightly such that they burst open at the vulva releasing their eggs. Then I permeabilize the animals by freeze cracking. My first problem is that the eggs then sometimes stick to the slides but the adults most times don't. How do I get the animals to stay on the slide? The second problem is that I have high nuclear background that fluoresces orange on the GFP filter, but has nothing to do with the fluorophore the secondary antibody is conjugated to. This background then interferes with staining visualized with FITC. What is this background? how do I get rid of it? I would appreciate any suggestions/ideas. Thanks in advance, Tali --- |
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| antibody , slides , staining |
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