>Dear list, I wonder if you can help me. I am trying to measure, using
>fluoresent dyes and confocal microscopy, production of reactive oxygen
>species,in particular peroxides in smooth muscle cells. I have tried to use
>5'6'carboxy-2'7'dichlorofluorescein-diacetate di(acetoxymethyl ester) without
>any success. The problem is that the dye activates and saturates as soon as the
>fluorescent light hits the cells. It has been used successfully in a limited
>number of expreiments using FACS analysis. Does any one out there have any
>experience in this field which could help me. Kind regards Sean.
>Email [log in to unmask]
 
Dear Sean,
 
        See our paper in Cancer Research {Akhlynina, T, V., Rosenkranz, A.
A., Jans, D. A. & Sobolev, A. S. (1995) "Insulin-mediated intracellular
targeting enhances the photodynamic activity of chlorin e6" Cancer Res. 55,
1014-1019
(1995)} - we used confocal microscopy and 2'7'dichlorofluorescIN-diacetate,
which is deacetylated in living cells; it is non-fluorescent but becomes
fluorescent (the fluorescEIN form) upon reaction with reactive oxygen
species. Light does NOT convert the fluorescin form to fluorescein.
 
        David
 
 
 
***********************************************
             Dr. David A. Jans
       Nuclear Signalling Laboratory
Division of Biochemistry & Molecular Biology
  John Curtin School of Medical Research
      Australian National University
GPO Box 334  Canberra  ACT  2601   AUSTRALIA
       Phone No.   <61> <6> 249 4188
         Fax No.   <61> <6> 249 0415
      e-mail No. [log in to unmask]
 
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