Clinical Applications of Flow Cytometry

Carleton C. Stewart and Sigrid J. Stewart

Laboratory of Flow Cytometry
Roswell Park Cancer Institute
Buffalo, NY

November 1995

Earn CEU's for this presentation through November 1997

Gating a Selected Population

Abstract:

A Cytologist or Pathologist uses a microscope to classify normal or abnormal tissue. This process can now be performed automatically by a flow cytometer and this new field has become known as automated cytology. All cells contain extracellular membrane or intracellular proteins that provide their functional activities. Each unique population of cells exhibit a repertoire of proteins that can be identified by staining cells with the right combination of fluorochrome conjugated antibodies. The flow cytometry generated pattern can then be interpreted for the definitive rapid diagnosis of non-malignant and malignant cells. In this presentation, the operation of a flow cytometer, the method for preparing cells and case studies to illustrate the technique will be presented.

LEVEL: BASIC

Learning Objectives

Upon completion of this presentation the participant will be able to do the following:

Describe the general application of the technique of flow cytometry in a clinical setting.
Describe a typical sample preparation procedure for analysis by flow cytometry.
List two interferences to Immunophenotyping by Flow Cytometry, and describe a method of compensation.
Describe the direct, physical measurements made by a flow cytometer used for subsequent analysis.
Define the data acquisition process used by all flow cytometers called, "list mode".
Given a typical univariate autofluorescence histogram, select the most probable marker position to distinguish between positive and negative cells.
Define the term "gating" as applied to Immunophenotyping by Flow Cytometry.
Given a typical bivariate histogram, label the quadrant markers.
Explain the difference in appearance between bivariate plots of samples from normal and ALL patients.
Distinguish between what the medical reimbursement community refers to as a clinical laboratory test (CLT) and a clinical anatomical test (CAT).

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