Re: CLINICAL question: bone marrow
BARRY GRIMES (bagrim1@pop.uky.edu)
Tue, 20 May 1997 22:12:14 -0400
Adrian,
A method that has been employed wih great success here at UK to help
visualize bone marrow as well as PB abnormalities is by gating using a
SSC-LOG (X) vs CD45 (Y) dot plot. By changing from linear to log and
lowering your SSC PMT by about 100 you can see the familiar "paw print" of
hematopoietic elements. As CD45 is employed (we use the Per-CP conjugated
form from BD to allow 3 color analysis on a FACScan) the cells are
separated nicely into quite distinct areas. As RBCs are CD45 negative they
will fall to the bottom of the plot with mature lymphocytes at the top.
Myelomonocytic elements will fall in between and be further separated by
the SSC-Log amplification. Blasts will segregate below the lymphs and to
the left of the monos and granulocyte precursors. With a little experience
abnormalities will often be instantly obvious in their deviation from the
norm.
The beauty of this method really becomes apparent when an abnormality is
less visually striking. Because of the nice separation it then becomes
quite easy to obtain pure gates and regions for further analysis within a
panel of MoAbs designed for lymphomas and leukemias.
Barry
Barry Grimes
Manager, Hematopoiesis FACS Laboratory
UKMC Hematopoiesis Center
800 Rose St.
Lexington,KY 40536-0093
606-323-8193
bagrim1@pop.uky.edu
CD-ROM Vol 3 was produced by Monica M. Shively and other 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: (765)-494-0757;
FAX(765) 494-0517;
Web
http://www.cyto.purdue.edu
, EMAIL
cdrom3@flowcyt.cyto.purdue.edu
