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The ability to operate a flow cytometer does not qualify an individual to
perform all types of flow cytometric tests in a clinical setting. Flow
cytometry in clinical diagnosis depends on the appropriate use of the
instrumentation with attendant quality control as well as proper selection of
reagents. Most importantly, there must be an extensive understanding of the
normal expression of antigens on the tissue being analyzed. The procedures
and approaches used for CD4 enumeration are not adequate for the analysis of
complex tissues such as bone marrow, lymph node or body fluids where normal
and aberrant cell populations are admixed. The major problem with flow
cytometry in a clinical setting is that it always gives data; an answer, a
number. Whether or not these data are relevant depends on the operation of
the laboratory, the correct procedures and the skill of the technical
individual assessing the data.
An underlying premise of the Consensus Conference on Flow Cytometry in
Leukemia and Lymphoma is that specimens for detection of hematopoietic
neoplasms must be run by people who are skilled in this complex analysis and
who have been properly trained in assessing the results. It is not proper to
accept clinical specimens or to perform clinical tests for which the
technologist or pathologist is not properly trained. The suggestion that
clinical bone marrow specimens are being submitted to be analyzed by someone
who is unskilled in this particular application is not only unacceptable but
unethical. Incumbent on the laboratory director and the technologist is the
knowledge of where their skills are limited and to refer a specimen to
another laboratory for more careful assessment.
Specifically, with regards to the analysis of bone marrow, training from
other individuals and laboratories is essential. A beginning is the annual
flow cytometry clinical course which has a large section on the analysis of
bone marrow specimens. Although possible to glean important tactics from
published manuscripts, the use of these on a daily basis is best understood
when observing the procedures in an operating laboratory. It is necessary to
spend time in an active flow cytometry laboratory which performs such tests
to develop a procedure manual, quality control procedures and gating
approaches which are difficult to create de novo.
Again, we sincerely apologize for our initial response.
Michael R. Loken, Ph.D.
Denise A. Wells, M.D.
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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