We need to find out the easy way for surgeons for store the tissue samples for us to do fluorescent in-situ hybridisation using DNA oligo probe later. Not sure whether tissue samples stored in RNAlater will affect fluorescent in-situ hybridisation.
Tissue should be rinsed in sterile saline or PBS, and snap frozen in OCT Compound, and stored at -80C.
Thanks Richard. One more question. The tissue snap frozen in OCT compound can be used for cryostat sectioning and mount on slide. People do fix tissue in 4% paraformaldehyde before doing fluorescent in-situ hybridisation. Do you fix the tissue after cryostat sectioning? Thanks again.
I work with rat brain tissue, which I snap freeze in isopentane/dry ice after dissecting out of the skull. After I section and mount the tissue slices on microscope slides, I'll store the slides at -80C until I'm ready to do the in situ, and I will fix the tissue in 4% paraformaldehyde before proceeding with ISH.
I would say the same as Susan.
-80°C until you are ready for in situ and 4% PFA.
Does anyone know why I should use 4% paraformaldehyde before proceeding with ISH?
I am working on flower buds, does these method also work well in plant?
4% paraformaldehyde does not cross-link proteins as extensively as glutaraldehyde. Unsure if the method is appropriate for plant tissue, however would suggest trying formaldehyde-alcohol-acidic acid (FAA)
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