Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Fura red should work. It can be as a single wavelength or ratiometric
dye. See:
Kurebayashi N, Harkins AB, Baylor SM. Use of fura red as an
intracellular calcium indicator in frog skeletal muscle fibers.
Biophys J. 1993 Jun;64(6):1934-60.
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=8369415
They used 420 and 480 nm excitation. Something I found out the hard way
when doing this is that you may need a secific "Fura Red" dichroic as a
lot of "~500LP" dichroics will pass, not reflect, the 420 nm excitation
light. This is also an issue with the ratiometric pericam proteins.
Compare:
Fura red:
http://www.chroma.com/products/products/filter_details.cfm?ID=71003http://www.omegafilters.com/front/curvomatic/spectra.php?part=XF100-2
FITC:
http://www.chroma.com/products/products/filter_details.cfm?ID=31001http://www.omegafilters.com/front/curvomatic/spectra.php?part=XF17
Also, we've had issues with Fura red AM loading mitochondria so keep the
concentration low.
Regards,
Tony
Kathy Spencer wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi All!
> Having difficulties with ligand autofluorescence in calcium
> imaging. We
> measure an auto-fluorescence peak around 460-570nm emission, and are trying
> Fura-2 and Rhod-2 calcium indicators. Our compound still interferes with
> the emission of these dyes. Can anyone recommend other calcium dyes we
> could try? We found that the Fluo dyes are right in the middle of our
> autofluorescence peak, but are there other dyes?
> Thanks!
> Kathy Spencer
>
>
>
> Kathy Spencer, Ph.D.
> Scientific Associate
> The Scripps Research Institute
> 10550 N. Torrey Pines Road
> ICND 202
> La Jolla, CA 92037
> 858-784-8437
--
Tony Collins Ph.D.
Facility Manager
Wright Cell Imaging Facility
Toronto Western Research Institute
13-407 McLaughlin Pavilion
399 Bathurst Street
Toronto M5T 2S8
tel. (416) 603 5367
