 |
 |
 |
 |
>Do any of you have experience with the vital stain Evans Blue?
>Specifically, someone wants to analyze cells stained with monoclonal:FITC
>and counterstained with Evans Blue, which fluoresces red, on our confocal
>laser scanning microscope. I'm concerned about emission spectrum overlap,
>but have been unable to locate information about the Evans Blue emission
>spectrum. Any information or leads will be appreciated.
Many years ago (pre PE and when people started worrying about HIV) our
diagnostic lab started to put Evans Blue in their cell fixative. This was
done so the flow operators would know the fixative had been added to the
sample! At this time they were just using single colour FITC labels and it
was established that the Evans Blue didn't cause any problems.
This fixative mixture became the `dept standard' cell fixative for FACS
work! When people started using PE, they couldn't get their two colour
samples to work at all. I ran some of this fixative through a fluorescence
spectro and as you can imagine it started fluorescing like crazy just below
600nm (if I remember correctly) and went on well into the red.
When the Evans Blue was left out of the fixative, naturally no more problems!
As I said this was a long time ago, but I could probably dig up the spectra
if you want it.
Leon Martin.
########################################################################
____
| | Dr. Leon Martin,
/ | _ Dept. of Pathology & Immunology,
| |-------> ( ) Monash University Medical School,
\ | ~ Commercial Rd., Prahran, Victoria,
|____| /| Australia, 3181.
__| |__ ___/ |
|_______|^^^^ --- e-mail: martin@cobra.path.monash.edu.au
| Voice: +61 3 276 2601
| Fax: +61 3 529 8359
--
########################################################################
|
|
|
|