We have the paraffin blocks from a lung of a dog suspected to have pneumocystis. Staining in H&E does not allow to detect fungal elements (cysts). Staining by Gomorri-Groccot is not available in our laboratory. Are there any alternative histological staining methods for visualization of pneumocystis? Or, are there any other diagnostic tools applicable for paraffin blocks?
A simple, quick and inexpensive option is to use optical brighteners (ex. Calcofluor white) if you have fluorescence microscopy with appropriate filter . It just takes dewaxing and then dyed.If you hold the material is a fantastic choice if you need more details feel free to contact. I can send photos of the characteristic cysts.
Dear Jose, thank you very much for your response! It sounds to be a great diagnostic tool for my case.
Gram-Weigert is also an option. See http://www.ral-diagnostics.fr/diagnostic/en/M-BAC/coloration/pc_vers_site/protocoles/Proto066_GramWeigertStainingforHistologicalSections_GB_V01.pdf
Dear Jose and Vladislav! Thank you so much for very useful information! I'll check both options and report my results.
You may use toluidine blue O
http://jcm.asm.org/content/26/4/774.full.pdf
http://www.rightdiagnosis.com/phil/images/5791.jpg
I see, Mona, thank you. To my knowledge, Gomorri-Groccot is also silver-based method.
Dear Roman if you would like to send us an unstained section to the Diagnostic Laboratories at the Royal Veterinary College, UK we can do the Grocott stain for you. If you could also send an H and E for comparison that would be helpful. Additionally I am interested in canine Pneumocystis so if you could also send a 5-7um section so that I can potentially look at the DNA I would very much appreciate that. Some of the antibodies used for detection in human laboratories may also cross react with canine pneumocystis, but not all of them will.
Dear Kate!
Thank you very much for your proposition. It is very helpful idea. I should check out the possibility how to send you the samples, and I let you know. It is very kind of you.
Hi Roman, according to Zeibig(1997) techniques such as monoclonal immunofluorescent staining have been proven helpful in identification.
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