Determination of nRBC in umbilical cord blood

From: Nick Greco <ngreco@clevelandcordblood.org>
Date: Thu Mar 27 2008 - 18:32:26 EDT
Cytometry Discussion group:



I am attempting to determine by flow the number of nucleated red blood cells
in cord blood by following the procedure of Tsuji T et al., Cytometry
37:291-301, 1999.  Briefly, the protocol uses EDTA-treated PB with
anti-CD45-FITC and PI (threshold FSC 52, FACSCalibur) with an acid hypotonic
lysis followed by a neutral hypertonic alkaline reagent.


I am using a BD Canto II with CPD-treated CB and we are following this
protocol but have been unable to see any nRBC in cord blood.


I appreciate any suggestions.



Nicholas J. Greco, Ph.D.

Director of Laboratory Operations
Cleveland Cord Blood Center
Suite 150

25001 Emery Road
Cleveland, OH 44128-0804

216-896-0384 (office)

216-896-0320 (fax)

440-488-9205 (cell)









image002.jpg
Received on Fri Mar 28 16:58:00 2008

This archive was generated by hypermail 2.1.8 : Wed Jan 31 2007 - 03:12:00 EST