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We are working with EGFP labeled cells in neural tissue from a transgenic mouse viewed live in oxygenated media using time lapse confocal microscopy on a Leica SP2. When we injure the cell, we would like to label the dead cell(s) with a
fluorescent stain. But I am not too thrilled with the dead cell labels, which bind nuclear DNA from broken cell membranes.
Propidium Iodide seems to stain, but is weakly excited by Argon lasers.
7-AAD may not pick up too well as a stain, but is better separated excitation laser wise.
Then there are the Molecular Probes SYTOX dyes, but they often require detergents.
Does anybody have a preferred dye/protocol to use for labeling dead cells in a live neural tissue?
Propidium iodide is also displaced by paraformaldehyde too.
Ethan.
Web:
http://go.usa.gov/cegV3
Ethan Cohen, Ph.D.
Div of Biomedical Physics, WO62 Rm 1204
Office of Science and Engineering Labs,
FDA Center for Devices and Radiological Health
White Oak Federal Res Ctr.
10903 New Hampshire Ave.
Silver Spring, MD 20993
Office: 301-796-2485
Lab:301-796-2762
Fax: 301-796-9927
Cell: 301-538-7544
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