I am trying to do an immunohistochemistry experiment in mouse aortic cross sections. I tried to stain with lamin but it didn't work. I have found in some papers that the researchers used proteinase before primary antibody treatment. I want to enlightened specifically about the staining of aortic sections and if I need to follow any extra measures to get the proper staining.
Regards.
H&E stain is a standard one: it can give you a general overview on your samples. But I guess if you started with lamin, you may want a more specific staining. Then CD31 and NG2 are also good options. But it depends on what you want to observe. And always remember to also put a reference stain, i.e. DAPI.
There are a lot more parameters to be considered, for example, you also have to decide which sectioning protocol to use, i.e. Paraffin, cryo-section, etc. Sometimes this makes a difference. The quality of primary antibody also matters.
In this case H/E stain would suffice. Detailed structure elements would all be well visible. If already started with lamin preferred counterstain would be Mayers Hematoxylin. Agreed on the reference stain as DAPI.
By using HE stain, trichrome stain, Verhoeff's or wigert;s elastica stains as conventional histological stains of paraffin sections.
reference for stain procedure:
S.K. Suvarna, C. Layton, Bancroft, J.D., (2013) Bancroft’s Theory and Practice of Histological Techniques, ed 7. Elsevier, Churchill Livingstone.
For immunohistochemistry you should follow the data sheet of antibody.You should select suitable antibody for the structure you need to observe or study it.
For example: 1–Prashanthi Menon, and Edward A. Fisher(2016)
Immunostaining of macrophages, endothelial cells and smooth muscle cells in the atherosclerotic mouse aorta.Methods. Mol Biol. 2015; 1339: 131-148
May be this article help you in staining procedure.
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