Hello,
Like the title says, I'm staining chicken sperm storage tubules with H & E to prepare for LCM. My tissues are frozen and sectioned via the cryostat with OCT. I just tried staining for the first time using the Histogene Frozen tissue staining kit by thermofisher, and my slides have a white chalk on them. I read online that this could be that the OCT has not left the slide. Is this correct? If this is the case, can I remove it with tap water after the slide thaws and before I start the dehydration process with 75% ethanol (as per the kit's instruction/protocol).
Hi Kyana
Put your slides at room temperature for 20-30 min, when the OCT will remove, use hydrophobic pen to make boundary around your section, then follow the H & E procedure. Do the washing step carefully. Don't let the solution come out the boundary.
Hi,
Thank you for your answer. So you're saying after thawing and marking, use an OCT solvent remover (like Formalin for example) + the tap or DI rinse before going into the staining protocol? Meetu Wadhwa
No, you don’t need any solvent for OCT removal. First just put at room temp, wait for sometime, OCT will automatically go out. If it still remains after 30 min, then wash with PBS.
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