Hi Kevin,
I use Sudan Black (from Humason’s Animal
Tissue Techniques). It is not fluorescent itself and works wonders on quenching
endogenous fluorescence, including from blood. The only issue is that the
staining is done in 70% alcohol to keep the dye soluble. So depending on your
other staining conditions, it might not work for you.
Good luck.
Helen McBride
Helen J. McBride, Ph.D.
Inflammation
Group
Amgen
One
Amgen Center Drive
Phone:
805-313-5476
Fax:
805-447-1982
Email:
From: Confocal
Microscopy List [mailto:[log in to unmask]] On Behalf Of Rosemary White
Sent: Thursday, July 12, 2007 2:19
PM
To: [log in to unmask]
Subject: Re: quenching of Alexa633
Hi Kevin,
You could try toluidine blue or crystal violet, though I’ve only used
these to quench stains fluorescing in the yellow-green. Or possibly basic
fuschin (from Schiff’s), which will fluoresce in the green-yellow but I
think is not fluorescent with red excitation. No experience with any of
them on animal cells....
cheers,
Rosemary
Dr Rosemary White
[log in to unmask]
CSIRO Plant Industry
ph.
02-6246 5475
GPO Box 1600
fax.
02-6246 5334
From: Kevin
Braeckmans <[log in to unmask]>
Reply-To: Confocal Microscopy List
<[log in to unmask]>
Date: Thu, 12 Jul 2007 16:04:45
+0200
To: [log in to unmask]
Subject: quenching of Alexa633
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear list,
We are using transferrin tagged with Alexa633 to label the clathrin-mediated
endocytosis pathway of living cells. We wanted to use trypan blue to quench the
fluorescence of the remaining Alexa-Tf on the cell membrane. However, this does
not seem to work, contrary to e.g. bodipy or fluorescein.
Do others also have found difficulties in quenching Alexa-dyes by tripan blue?
Does someone know an alternative quencher which might work?
Thanks for any suggestions,
Kevin
Kevin Braeckmans, Ph.D.
Lab. General Biochemistry and Physical Pharmacy
Harelbekestraat 72
9000
Tel: +32 (0)9 264.80.78
Fax: +32 (0)9 264.81.89