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Pedro Camello |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi all, I-m trying to record, in living cells, mitochondrial FAD autofluorescence and TMRM. I use a Nikon A1, using (exc/em); FAD 476/505-530, and TMRM 561/590-620 or a long-pass filter. While exciting with the 561 laser I´m finding some fluorescence at the lower emission window (505-530). Unexpected to me. I´ve been unable to locate in the web a FAD fluorescence spectrum. My questions: Could any A1 spectral user suggest me how to set the optimal conditions to differentiate FAD and TMRM fluorescence? Could anybody supply me with a FAD fluorescence spectrum? Thanks in advance |
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George McNamara |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Pedro, Fig 4 and 5 of Benson 1979 J Histochem Cytochem (www.jhc.org) has em spectra of riboflavin, FAD, FMN (Fig 5: all three practically identical). Figure 3 of Aubin 1979 J Histochem Cytochem has riboflavin ex and em spectra - will be similar to FAD. Fig 2 of Biophysical J has em spectrally unmixed components that authors claim are flavin-ETF, free FAD, flavin-LipDH. See also Maeda-Yorita and Aki 1984 J Biochem 96: 683-690. Enjoy, George On 2/9/2011 2:52 AM, Pedro J Camello wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi all, > > I-m trying to record, in living cells, mitochondrial FAD autofluorescence > and TMRM. > > I use a Nikon A1, using (exc/em); FAD 476/505-530, and TMRM 561/590-620 or a > long-pass filter. > > While exciting with the 561 laser I´m finding some fluorescence at the lower > emission window (505-530). Unexpected to me. > I´ve been unable to locate in the web a FAD fluorescence spectrum. > > My questions: > > Could any A1 spectral user suggest me how to set the optimal conditions to > differentiate FAD and TMRM fluorescence? > > Could anybody supply me with a FAD fluorescence spectrum? > > Thanks in advance > > ... [show rest of quote] -- George McNamara, PhD Analytical Imaging Core Facility University of Miami |
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