Hello there,
I want to snap-freeze mouse kidneys for immunofluorescence using isopentane surrounded by liquid nitrogen (1/4 of kidney for 1 min and then rapidly in -80°C freezer).
Is there any advantage or disadvantage - concerning freezing process, storage or sectioning - to cover the tisse with OCT (e.g. in a cryomold) before placing it in isopentane or can you simply embedd it in OCT using dry ice one day before sectioning?
Thanks.
Dear Konstantin,
we usually wrap the tissue in aluminium foil and than put it into isopentane, cooled by liquid nitrogen. In our experience covering with OCT before freezing has
no benefits.
Best regards, Daniela
Have you considered using the procedure w/o OCT. Perhaps you could snap freeze as you stated, keep the tissue in -80. Only in al later stadium we use OCT to glue the specimen to the cryotome holder. Keep as much tissue free. We cut 4 um sections and catch them on lysine coated slides
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