Flow Cytometry and Sorting
Facility
Director: G. Larry Gartland, PhD
Department/Center
Association: Division of Developmental
and Clinical Immunology
Established: 1980
The
Flow Cytometry and Cell-Sorting Facility was
established in 1980 by what is now the Division of Developmental and Clinical
Immunology. Its aim was to provide
support for studies of lymphocyte development and function by furnishing a
physical means of identifying and isolating lymphocyte subsets. Since then, the facility has specialized in
the analysis and separation of various lymphoid- and myeloid-lineage cells and
stem cells.
Facility Description
The
facility has two cell sorters: a 2-beam FACStar Plus (with 488nm and UV
excitation) and a high-speed, 3-laser MoFlo (with argon, krypton, and
tunable-dye lasers). These instruments
are used to analyze and sort cells on the basis of several light-scatter
measurements and, in the case of MoFlo, of up to eight fluorescence
signals. MoFlo incorporates provisions
for sorting cells into wells of arbitrary rectangular plates and for separating
up to four subpopulations of cells simultaneously.
The
Division also maintains three flow cytometers, a FACScan,
FACSCalibur, and Cyan, which are used by individual
investigators for fluorescence analyses of cells. These instruments can measure three, four,
and seven fluorescence labels per cell, respectively. All instrumentation is housed on the fourth floor
of the
Research Information
The
facility is used extensively by members of the Division for a variety of
analytical and sorting procedures.
Current use within the Division averages nearly 200 hours per month.
Analyses. Most
analytical procedures are carried out by investigators themselves using the FACScan, FACSCalibur, or Cyan. Typical applications include hybridoma screening, cell immunophenotyping, multi-color
subpopulation analyses, cell cycle analyses, and calcium mobilization
measurements.
Cell
Sorting. Cell sorting is performed daily and
accommodates a range of experimental designs depending on the cell type, the
number of fluorochromes used, and the number of cells collected. Some examples are single cell cloning, mouse
stem cell sorting (tens of thousands), lymphoid subpopulation separations (tens
of millions), and lamprey lymphocyte-like cell isolation (hundreds of
millions).
Although
the facility is not a university-wide core facility, its personnel try to
assist researchers from outside the Division whenever the schedule allows.
Core Director: G.
Larry Gartland, PhD
Email: glarryg@uab.edu
Phone:
205-934-0331
Approved by: G.
Larry Gartland, PhD, Director
Date: March 22,
2008
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