Hello Researchers,
I have been trying to do a BODIPY stain of lipid droplets in THP-1 cells without luck. The images are fuzzy and blurry at 63x magnification on confocal microscope. I had initially done this in an U well slide and I got good images. I started getting blurry images when I plated the cells on conventional cover slips to put on slide
If the paraformaldehyde solution is buffered, it can be stored refrigerated for a couple weeks. pH change and breakdown formic acid will occur faster at room temperature. "If the solution is unbuffered, the pH may drop below 6.2-6.4. In this case, byproducts of paraformaldehyde decomposition (formic acid, gaseous formaldehyde, etc.) may form and give rise to formalin pigment. The pigment must be removed from this solution to avoid false staining." https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/270/236/p1213pis.pdf
I wonder if the mounting of the coverslip grown cells compressed them, distorting the lipid droplets.
Hi! Also eg glycerol containing mounting medium such as mowiol can affect LDs, induce fusion etc. I would suggest mounting samples in pbs if possible.
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