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Carol, based on the information you gave you can use either FM-123 or
Propidum iodide for counter staining membranes. You can try the standard
FDA (Fluoroscine diacetate) to both assess the live cells as well as the
cytoplasm. These dyes works best for live cells. If you use fixed cells
then we need to think something else.
Shiv

Mayandi Sivaguru, Ph.D.,
Associate Director
Molecular Cytology Core Facility
Molecular Biology Program
2, Tucker Hall
University of Missouri
Columbia, MO 65211-7400

Voice: 1-573-882-4895
Fax: 1-573-882-0123

www.biotech.missouri.edu/mcc/


At 04:11 PM 12/13/02 -0500, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hello,
>I am helping someone determine whether their protein is on the
>membrane, inside the cell, or both.  the living cells (I don't
>remember what kind they are) are grown on cover slips and are very
>flat.  It is difficult to tell.  they are thinking of counterstaining
>with a cytoplasmic dye.  Several come to mind, like rhodamine 123 for
>mitochondria, or a Cell Tracker dye.  conversely one could try to
>stain the cell membrane.
>Does anyone have any suggestions?  Would it be OK to use cells that
>are rounding up? They are the brightest but I'm worried their
>membranes may be compromised.
>Thanks in advance.
>Carol
>--
><><><><><><><><><><><><><><><><><><><><><><>
>Carol Bayles
>Microscopy,  Imaging & Fluorimetry (MIF)
>Bioresource Center
>160 Biotech Bldg
>www.brc.cornell.edu
>
>Confocal and Multiphoton Microscopy
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>
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