Posted by
Rietdorf, Jens on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Quantifying-fluorescence-help-tp593618p593626.html
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalDear Maria,
(no commercial interest) Invitrogen sells a product called InSpeck, ie
beads with a calibrated intensity of labelling. They come in different
intensities, like 100%, 50% 25% usf. If you include those in all your
preparations you can normalise your signal to the beads, which may be
sufficient to have a rough idea of the relative brightness of your
labelling.
Cheers, jens
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On
Behalf Of Maria Mazzillo
Sent: Freitag, 18. Juli 2008 15:26
To:
[hidden email]
Subject: Quantifying fluorescence help
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalI am a PhD student at Auburn University just getting started into my
thesis work. I am looking for a reliable method for quantifying
fluorescence.
My project deals with marine dinoflagellates (zooxanthellae) that reside
intracellularly in hosts (usually cnidarians). I am working with
cultures or isolates of only the dinoflagellates for my confocal work.
I have an antibody that was created against the surface secretions of
mucilage (secreted as part of a daily cycle by the alga) from one strain
of zooxanthellae.
I am attempting to use this antibody to label various strains to
identify differences in mucilage between them. Thus far, I have seen
that the strain the antibody was created against labels around the cell
fairly brightly. Most samples either show this or a complete lack of
labeling. However, a few samples show a faint fluorescence lifted off
the cell surface. I would like to be able to quantify this fluorescence
in comparison to either the control strain that the antibody was created
against or against a known fluorescence.
Any help on ideas for this, or places to look for ideas would be greatly
appreciated.
Thanks!
Maria Mazzillo