Hello,
I recently stained my cells with an Arf6 antibody. The healthy control looks pretty normal, while the disease group cells have abnormally large black structures in the cell body. Has anyone experienced anything similar? Are those holes, vesicles, or anything else??? For further info, those cells are destined to die, so they might as well be disintegrating at the fixation time. Still, I am curious about what I saw.
Elif Bayraktar , 'digestion', 'lipidic' or 'enzymatic' 'vacuoles' ["...ARF genes...play a role in vesicular trafficking and as activators of ....phospholipase D."] come to my mind immediately. Unfortunately I haven't seen such cells in culture stained by Arf6 (just for the record and for correct 'RG-archive search phrase" reasons: Arf6= ADP-ribosylation factor 6) Antibody.....
But: I don't think it were 'disintegrating, previously nuclear' bodies....
To get proof of the nature of those "holes" or "black structures" (as you named them), in an attempt of correlative light microscopy, proper chemical fixation and processing at least into paraffin wax (if you have options for and would think of cryofixing / cryoprocessing, staining, etc....you might try...but I'd guess, the results wouldn't reveal the "real nature" of the morphological conspicuities) seems mandatory.... To achieve optimal results IMHO one would perform classical specimen preparation for TEM-observation on the ultrastructural scale...
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