Re: Indo-1/[Ca++]i calibration

From: Larry Arnold (lwarma@med.unc.edu)
Date: Wed Nov 28 2001 - 09:41:08 EST


I should have noted that the cells are alive at the indo-1 loading and load
fine.  The problem is happening after the cells are treated with ionomycin,
nigericin et al. and placed in the Ca++ buffers.  I'm assuming that then
the membrane becomes permeable and the indo-1 leaks out.

Larry

At 09:15 AM 11/28/2001, you wrote:
>Larry,
>         The calibration can't be done with cells that are dead since the indo
>1 ester won't be hydrolyzed. We do our calibrations with live cells, get
>readings before and after ionomycin (no other permeabilizers) and have no
>problems.
>         Liz

Larry W. Arnold, Ph.D.
Associate Professor
Director, Flow Cytometry Facility
Department of Microbiology and Immunology
Lineberger Comprehensive Cancer Center
CB# 7290
University of North Carolina at Chapel Hill
Chapel Hill, NC 27599
Phone: 919-966-1530
FAX: 919-962-8103



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