Hi everyone
Any possibility to do Immunohistochemistry (IHC) on retrospective(old) Bone Marrow/Peripheral leukemia(AML) smears slides stained already with Giemsa/Wright
I want know about the possibility of doing the Immunohistochemistry (IHC) on retrospective(old) Bone Marrow/Peripheral leukemia(AML) smears slides made 2-3 years back.
The slides are old and stored and already stained with Wright's/Giemsa and Wright-Giemsa stains
So, i would like to know is it possible to IHC for any protein expression on it. I want to the see expression of proteins ( B catenin, APC, and Axin).
How feasible is it to do and how would be its results, keeping in view the old and stained slides??
Please share your experience about this and the related articles or literature.
Regards
According to my limited knowledge in this area I most think that its impossible,well its possible but the results are not going to be valid.
The cells are already fixed and stained, don't think that it's wise to use these slides.
I normally have some spare unstained slide of my samples.
Johan, even if Mohsin had unstained slides from the same experiment, wouldn't the 2-3 years of storage make staining impossible? What was the longest you have waited between cutting and successful staining? I'm just curious, for future referenece.
ah i thought they were younger :) now i see the 3 years,
I got experience with superresolution with slides from 6 months back and they were fine, now i'm doing stainings on BM and PB slides from the beginning of 2014 they look good as well.
further then 1 year i would avoid, but if you have good controls you can always check.
the big message is, avoid long storage, avoid freezing best will still me fresh samples but, due to availability sometimes you will have to do with what you got!
and always take a good control (cell line) and secondary ab control.
Hi
Actually, i got an idea from the already posted same query on Rech gate, some have done it,........donot exactly their conditions of slides
https://www.researchgate.net/post/Is_it_possible_to_do_immunohistochemistry_detection_on_bone_marrow_smears
further suggestions and guidance is welcomed
Use microwave techniques: see Boon and Kok Microwave cookbook for microscopists or the newer version: Microwaves for the Art of microscopy. See also the special issue J neurosci methods 55/2/1994, old but also look for antigen retrieval: see Shi et al J Histochem Cytochem 39: 741-748, 1991; Cattoretti 1992 Histochem J 24:594; Momose et al., 1993 Appl Immunohistochem 1:97-82; Suurmeyer and Boon (1993)Eur J Morphol 31:144-150 and J neurochem 30:361-371.
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