Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

And maybe you do not even need an acceptor, because all of what is
regarded as 'autofluorescence' is excited and generates damage. Remember
how easy it is to kill an unlabeled living specimen just by strong or
long illumination. I dont know about Poly(allylamine hydrochloride)
though, maybe its worthwhile checking if there is 'autofluorescence'
from it?

Iain Johnson schrieb:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> This doesn't surprise me at all.  The acceptor photobleaching technique
> is not necessarily selective for the acceptor.  Excitation of the
> acceptor photosensitizes generation of singlet oxygen that then reacts
> quite indiscriminately with dyes, proteins, nucleic acids or any other
> reactive target in its path. Since the diffusive range of singlet oxygen
> in living cells is about 50 nm and therefore of the same order as the
> donor-acceptor proximity required for FRET, it is quite possible for
> singlet oxygen generated by excitation of the acceptor to bleach the
> donor.  This is one reason why FLIM detection of FRET, which does not
> suffer from this vunerability, is a superior technique to "acceptor"
> photobleaching.
>
>
> Alberto Diaspro wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Dear friends,
>> in order to check some possible FRET between FITC and ALEXA 555, when
>> using bleaching of acceptor as method, we found that 543 nm bleached
>> FITC as well.
>> Did you ever experienced such a behaviour?
>> Alby
>>
>>
>> ------------------------------------------------------------------------
>> ---------------------------
>> Alberto Diaspro, Department of Physics, University of Genoa
>> Via Dodecaneso 33, 16146 Genova, Italy
>> facsimile +39-010314218 - voice +39-0103536426/480/309
>> URL: http://www.lambs.it
>> http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html
>> ------------------------------------------------------------------------
>> --------------------------
>
>
>

--

Dr. Jens Rietdorf
EMBL, Meyerhofstr.1,
D-69117 Heidelberg,
Cell Biology/Biophysics Program,
Advanced Light Microscopy Facility
Phone ++49(0)6221 387-467 FAX-306
http://www.embl.de/almf