http://confocal-microscopy-list.275.s1.nabble.com/Concentrating-bacteria-cells-for-microscope-visualization-tp6519025p6526033.html
Lucifer yellow reaches yeast cell (S. cerevisiae) vacuoles by fluid-phase
endocytosis in an hour. Might work with mammalian cells too.
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>
> Hi all,
> This is not a "confocal" question but rather a cell bio/microscopy
> related one. Nevertheless, I am hoping someone will be able to answer.
>
> I co-teach a cell biology lab course where students learn how to use
> conventional epi fluorescence and video microscopy. We have a variety of
> "cells in action" labs for them to do, one of these being endocytosis. As
> markers, e have used labeled-transferrin and also DiI labeled LDL, with the
> idea being that the transferrin is recycled back to the plasma membrane
> whereas the LDL moves into lysosomes. While the transferrin labeling seems
> to work more or less as expected, results with the DiI LDL are erratic.
> Sometimes there is no labeling at all, and we never have seen what looks
> like deep internal labeling in putative lysosomes.
>
> We plan to troubleshoot later this summer but as none of us have
> studied endocytosis, we have no first hand experience. I am wondering if
> anyone out there knows of a tried and true endocytosis cell bio lab? Or
> perhaps knows about some tricks for handling DiI LDL. Or perhaps it is a
> matter of a good cell line -- we work typically with a 3T3 fibroblasts or
> LLCPK epithelial cells (with equally dismal results for the DiI LDL
> endocytosis).
>
> Thanks in advance for any suggestions.
>
> As ever
> Tobias Baskin
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