I am investigating the effect of an epileptic drug on pregnant rats and their embryos, so I would like to investigate whether or not necrosis occurs in the liver, kidneys and brain tissues.
I do not know if this could be useful for you but I will tell you
In some necrotizing melanomas, it has been described that the necrotic areas are completely negative to cytokeratin AE1/AE3, to CD31, chromogranin, and SMA. The borders of the necrotic area could be marked with antibodies anti-CD45. The reaction against CD45 will be predominantly present at borders between the viable and necrotic areas, and the positive cells will be morphologically consistent with infiltrating lymphocytes. Furthermore, similar immunoreactions were also observed with antibodies anti-CD68, which highlight the infiltrating macrophage population.
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Lactate dehydrogenase estimation is suggested as a marker of cell injury. Membrane damage & leakage of intracellular contents are hallmarks of cell death.
I also happened to read about RIPK1 ( Receptor interacting serine/threonine protein kinase1 ) which is thought to be involved in enzymatic activation of necroptosis. RIPK1 antibody may be used for IMMUNOHISTOCHEMICAL analysis. However ,I feel that morphological analysis of H& E sections would be the initial step in the assessment of any pattern of cell death, followed by Immunohistochemistry. Electron microscopy would give greater clarity of cellular membrane alterations & sub cellular alterations, however this depends on the scope of the research project .
The immunohistochemical study of the coagulative necrosis using formalin-fixed and paraffin-embedded tissues,demonstrated that cytokeratins (AEI/AE3 and CAM5.2) and carcinoembryonic antigen (CEA) were well preserved. However, compared with viable tumor tissues, only a few tumor cells were positive for epithelial membrane antigen.
Dear Marcello, thank you very much for your answer. However, I hoped to find a marker that distinguish necrotic cells themselves like the case with caspase3 or TUNEL used to distinguish apoptotic cells.
Dear Dr Bunglavil, thank you very much for your answer, it was very helpful. I agree with you that H&E is the main step in distinguishing necrotic cells, as well as Electron microscopy, however my research project needs immunohistochemical technique application.
Dear Ahmed, thank you very much for you contribution , however I would like to know if these markers give positive results with non carcinogenic or tumor necrotic cells or not.
As far as the current situation can tell, there is an evident lack of reliable necrotic markers. You need to start with examining the histological slides then go through the EM investigation which will give a clear picture for the process of necrosis within the tissue under consideration. It seems that there are considerable efforts with regard to determine a reliable necrotic marker, hopefully see the light in the not so far future.
Immunostaining for complement C4d identifies necrotic heart muscle cells in FFPE endomyocardial biopsies from heart transplant recipients - it identifies complement activation which, theoretically at least, does not occur in apoptosis.
Hi all,
I am suffering to distinguish apoptosis from necrosis in Newcastle disease virus-infected birds and we are dealing with a newly emerged virus that behaves differently in different tissues such Bursa of Fabricius and spleen in infected birds,
My supervisor suggets DNA degradation assay but it has lot of false possitive in FFPE tissues,
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