RE: fix first stain later for surface antigens

From: Dr. Debbie Glencross (debbieg@mail.saimr.wits.ac.za)
Date: Fri Nov 29 2002 - 04:17:11 EST


Dear All,
We have used Transfix for transportation of samples from Johannesburg to
London, Royal Free Hospital, with great results (Glencross et al, Cytometry
2002; 50 (2); 69-77) and Jani et al, 2002(listed below) . Transfix enables
fixation of samples so that they can be tested up to 10 days after
venesection, without loss of flow scatter or antigen expression (basically
they remain the samples retain the characteristics of fresh samples up to 10
days after collection).

Our angle was aimed at showing  (1) that Transfix fixation facilitated
testing without loss of cell count or antigen expression (enabling us to do
comparative testing) in sites thousands of km apart, and further (2)
demonstrated our initial aim for use of such fixatives in resource poor
settings, to increase accessibility of testing in third world countries. In
other words, Transfix could facilitate transport of samples from remote
areas, where sample fixation could be performed in a laboratory with very
basic pathology facilities, to more high tech referral centre for testing
(e.g. to facilitate a flow CD4 T cell counts rather than advocate the use of
lesser technology like manual bead counting) (Jani et al, Journal of
Immunological Methods 2002; 257: 145-154.)
See also:
Barnett D, Granger V, Pockley AG, Saxton JM, Storie I., Whitby L and Reilly
JT.  TransFix: A Sample Stabilising Fluid for use in Cellular Hematology and
Immunology. Cytometry (Communications in Cytometry) 1998; 38:88 (abstract).
Shafaie A, Bergeron M, Phaneuf S, Mandy F, Granger V, Reilly JT and Barnett
D.  A Robust Whole Blood Preparation for Absolute Counting. Cytometry 2000;
Suppl. 10; 145.
Shafaie A, Bergeron M, Mandy F, Granger V, Reilly JT and Barnett D. (2000)
TransFixed whole blood may be a QC solution for the single platform cell
enumeration. Cytometry, Suppl. 10: 146.

Contact David Barnett at UK NEQAS and I am sure that he will gladly help you
out with some Transfix supplies.
D.Barnett@sheffield.ac.uk

Debbie Glencross


-----Original Message-----
From: Fishbaugh, Justin [mailto:justin-fishbaugh@uiowa.edu]
Sent: 26 November 2002 12:26
To: cyto-inbox
Subject: fix first stain later for surface antigens



Greetings,

We have an investigator that needs the ability to fix human blood and stain
later for surface and possibly intracellular antigens.   A search of Medline
came up with Jani et al, "Affordable CD4+ T cell counts by flow cytometry
II. The use of fixed whole blood in resource-poor settings," Journal of
Immunological Methods, Volume 257, Issues 1-2, Pages 145-154.  This article
talks about using a fixative called TransFix made by UK NEQAS before
staining.   A search of the UK NEQAS website did not find anything
referencing TransFix.  Does anyone have experience with this stuff or have
some other magic potion that would do the trick?  Thanks.

Justin Fishbaugh
Technical Director
Flow Cytometry Facility
Holden Comprehensive Cancer Center
University of Iowa
48 EMRB
Iowa City, IA  52242
(319) 335-8103 Tel
(319) 335-8049 Fax
http://www.medicine.uiowa.edu/flowcytometry/



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