Re: endocytosis for cell biology lab

Posted by G. Esteban Fernandez on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Concentrating-bacteria-cells-for-microscope-visualization-tp6519025p6526033.html

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Lucifer yellow reaches yeast cell (S. cerevisiae) vacuoles by fluid-phase
endocytosis in an hour.  Might work with mammalian cells too.
http://jcs.biologists.org/content/117/14/2983/F6.expansion.html

-Esteban

On Tue, Jun 28, 2011 at 11:25 AM, Tobias Baskin <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy>
> *****
>
> 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
> --
>      _      ____          __   ____
>     /  \   /          / \    /   \ \        Tobias I. Baskin
>    /   /  /          /   \   \      \         Biology Department
>   /_ /   __      /__ \   \       \__    611 N. Pleasant St.
>  /      /          /       \   \       \        University of Massachusetts
>  /      /          /         \   \       \          Amherst, MA, 01003
> /      / ___   /           \   \__/  \ ____
> www.bio.umass.edu/biology/**baskin<http://www.bio.umass.edu/biology/baskin>
> Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
>