Re: Thiazole Orange

TERRY HOY (WHGTH@CF.AC.UK)
Thu, 19 Jan 1995 10:06:46 GMT

> I am currently working with the dye thiazole orange, and so far the research
> project has been limited to a FACScan. However, we'd like to move on to the
> next step, which would involve sorting on a B-D FACS440. I have found a few
> articles in the literature where individuals have used this dye on a FACSTAR
> PLUS. Has anyone ever used this dye on a sorter? I thought it was in the same
> chemical family as acridine orange, and I know that acridine orange gets
> embedded in the tygon tubing and results in tubing replacement. The FACS440
> has even more fluidic tubing than the FACSTAR PLUS. I was planning on just
> running 10% bleach through the fluidics after the sort. Is this adequate for
> removing all residual dye?
>
> The working concentration we are using is 1 ug/ml. Thanks so much for any
> comments on this topic.
>
> Marilyn Dietrich
> Louisiana State University

In the past we have sorted reticulocutes stained with thiazole orange
on a FACS440. There were similar problems to those you are aware of
with acridine orange.

We reserved a complete nozzle holder, nozzle and sample line for
thiazole orange work as we found this only took a few minuets to
swap over and guaranteed that no dye was lurking in the system.

Our decontamination procedure for thiazole orange and acridine
orange on our FACScan and FACS440 is 10 mins. ethanol to dissolve
any residual dye and 10 mins. 7X to remove any cellular debris.
We do not use any bleach.

Hope this information is useful to you.

Terry Hoy
Department of Haematology
University of Wales College of Medicine
CARDIFF CF4 4XN
UK

Home Page Table of Contents Sponsors Web Sites
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu