We have a protocol to detect a certain protein on bone marrow section, could this be adapted to bone marrow smears? What are the problems related to that? Any protocol is very welcome!
I have done bone marrow smears with Wright Giemsa, and Prussian Blue/Iron staining. I have also done Acridine Orange staining on smears but no IHC yet. I can think of no problem if the cells are fixated well and make it through antigen retrieval (~100 temp/1 hr). I certainly would never suggest microwave protocols for antigen retrieval in BM, I have yet to see a good one. If you have multiple slides per sample (I know, I know, not always possible), I would certainly try. I believe it would work, which antibody are you using??
thanks! we are using a regular antibody of a protein we believe we could detect on several precursors, but it's definitely not a routine test.
Thanks for the tips I will take them into account!
bene
We have done IHC on blood films and tissue smears on glass slides so bone marrow cytology should be the same. See: Chang, Li-Wen. Development of Molecular Diagnostic Tests for Inclusion Body Disease in Boid Snakes. 2012. ProQuest, Ann Arbor, MI, USA through University of Florida, Gainesville, FL, USA.
http://ufdc.ufl.edu/UFE0045011/00001. See Chapter 4. Immuno-based diagnostic tests for screening IBD.
I have done IHC on tracheal wash samples of foals (See: http://www.sciencedirect.com/science/article/pii/S0021997510003440).
You can also prepare cell blocks, it is easier with blood.
we have done immunocytochemistry ( immunoperoxidase) on BM & blood samples. This need cytospin preparation to have adequate number of cells in small area on slide. It does not require antigen retrieval because it is does not go through paraffin embedded steps. so we skipped this step. Results are very good.
The bone marrow can do like the blood smear., air dry and fix in 4c acetone for3-5min or in methanol. You can keep those slide in 4c for a week before ICC.
It is not necessary to use antigen retrieva,l therefore the antibody could be more diluted and reduced the incubation time.
You can have very good results using polylisinated slides, and dry smears for 1 h beforre staining.
We are doing IHC on cytology and FNAC tissue smears by fixing in cold acetone and the results are good;hence it should not be a problem on bone marrow smears
we do (or did) IHC on cerebrospinal fluid (CSF) and tumor samples (smears). IHC works well for both with a variety of antibodies. However, different antibodies may require different fixation for optimal staining results. We usually use isopropanol at room temperature for 10min. Some antibodies work better when slides are just air dried or fixed at -20°C in aceton or fixed at -20°C in aceton/methanol. Sometimes additional formalin fixation (5 min at room temperature are long enough) of the slides may also be helpfull. If isopropanol alone doesn't work well, we just try another fixation.
Hi everyone
I want know about the possibility of doing the Immunohistochemistry (IHC) on retrospective(old) Bone Marrow/Peripheral leukemia(AML) smears 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 will its results, keeping in view the old and stained slides??
Please share your experience about this and the related articles or literature.
Regards
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