Hi everyone,
I recently did a TEM on fibroblast cells and noticed that the cells, including control (normal and untreated)cells appear torn (or massive vacoules). The fixing and processing of the cell pellet was performed by a lab that offers these services. I have attached a figure for you to look at. Can anyone tell me if this is normal for skin fibroblast cells or was there an error in the processing? If so, what would you recommend doing?
Thank you
Hi Paalini,
Swelling of the endocytic/exocytic vesicles often happens in chemically fixed samples (the 'vacuoles' you mentioned). The tubular extrusions do exist on fibroblasts but can also be caused by the processing. Did you grow the cells in suspension or as adherend cells? In the latter case trypsinizing or scraping the cells to get a suspension before fixation can also cause these effects.
in that case it is better to do the fixation first and then scrape the cells in 1% low gelling point agarose and pellet the cells by centrifugation. after solidification of the agarose the pellet can be cut in convenient pieces and further processed for resin embedding.
best,
Rob