I have immunohistochemistry slides of human subcutaneous fat biopsy, with skin attached. Most of the structures are self-evident - skin, adipocytes, microvessels in transverse section (revealed by staining with vWF) and collagen; but present in all sections there are isolated small multi-cellular structures which are a mystery to me (see attached image). vWF staining on sequential slides shows these cellular structures not to be vascular. I welcome suggestions as to what they might be.
Good day, Christopher Kelsall
You are right at this figure group of microvessels situated in the center? but they have perivascular oedema (red arrows). If you look at the blue arrows at the top of the figure you will see connective tissue fibers, but at the bottom of the figure, it is homogenous due to disorganization of connective tissue (violet arrows). It seems to me that perivascular expression of vWF caused by plasmorragia and disorganization of connective tissue.
P.S.I didn`t find fat tissue at the presented figure
Best regards,
Dmitry
Hi Dmitry, thanks for your answer. I should perhaps have clarified that the section was prepared from frozen tissue, hence the tissue disorganization; I should also have clarified that this section was not stained with vWF, but that the vWF staining which was performed on a sequential section did not show these cellular bodies to be microvessels, although I agree that's what they look like. I might have chosen a better slide to illustrate the conundrum. There is no fat (or skin) observable at this scale; I felt that a x10 image would not have shown the cellular bodies of interest clearly enough.
Good day, Christopher Kelsall
Thank you for your answer. I will be grateful if you also add micro photo at magnification x400 of DAB+ cells.
Best regards,
Dmitry
Hi Dmitry,
I lack the microscope equipment to do x400; I can do x100, but the images are not of good quality because of unavoidable vibration in the lab, and limited illumination of the specimen.
Hi Dmitry,
I have added some more images which better illustrate my original question. You can see where the vWF staining is, which does not coincide with the same cellular structures on sequential slides stained for GLP1R.
Dear Christopher,
Could you elaborate abit more on your IHC staining procedure.
Thank you,
Ivan
Hi Ivan,
I didn't do the IHC, it was done by a professional pathology service. Sections were cut from specimens frozen in isopentane at -70oC and subsequently fixed with paraffin. This was necessary because the IHC was not planned prior to archiving of the samples. Consequently, there will be freezing artifacts in the sections. All sections were stained with primary antibodies (GLP1R or vWF) and DAB-conjugated secondary antibodies. Positive control slides (not shown here) confirmed the viability of the primary antibodies at the concentrations used.
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