Is there a difference between immunostaining for 4% PFA-fixed tissue and frozen tissue?
I would appreciate any guidance on immunostaining for frozen tissue, as this is my first time working with it!
Hello, in our laboratory for frozen tissue we are previously fixing tissues (brain) in pfarmaldehyde during transcardial perfusion and then in 30% sucrose + paraldehyde for a month. After that brains freeze in special compound (O.C.T. compound, by Sakura) and slice it using criotome 15-30 mkm. Very important find proper antibody, wich is can be used for frozen tissues, becouse sometimes another AB for parrafine or free floating slices can't work on frozen
Frozen section could also mean non fixed tissue shock or quick frozen in Isopentan over liquid nitrogen or dry ice. It depends on your kind of Immunohistochemistry and the used Antibodies. I would recommand to use Antibodies for PFA or Formalin fixed tissue as Kristina has discribed it. If you are using thick sections that means thicker than 15 micrometer then you should use the free floating method. Because you will get more intensive and homogeneous results. If you are working with sections thinner than 15 micrometers the slide method is highly recommended.
Hello Kristina Smirnova, Thank you for your suggestion about choosing the Ab! We usually do the same thing to fix the tissue in the lab.
Hello Ute Neubacher , Thank you for your advice! They said the frozen tissue was in the dry ice.
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