method to anchor non-adherant cells for Calcium imaging?

James Tsuruta jtsuruta at med.unc.edu
Tue Dec 16 17:46:18 EST 1997


Help! We are attempting to image calcium fluxes in spermatogonia after
loading them with FURA-2 and treating them with various growth factors.  

The problem:  spermatogonia are non-adherant cells and wash off of our
coverslips making subsequent manipulations problematic :(

We've tried/are trying to use coverslips coated with:

poly d-lysine
Collagen I or IV
laminin
laminin + collagen IV

These methods do not anchor our spermatogonia very tightly.  Does anyone
have a protocol for anchoring non-adherant cells to coverslips or
TC-dishes? 

Thanks in Advance,

Jim Tsuruta
Laboratories for Reproductive Biology
University of North Carolina at Chapel Hill



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