Washington State University Flow Cytometry Laboratory
The Flow Cytometry Lab is housed in the Animal
Disease Biotechnology Facility (ADBF) Room 4075 and is operated
by the WSU College of Veterinary Medicine.
The FC Lab has two analytical flow cytometers (FACS Calibur and
FACSort) and one flow cytometer with sorting capability (FACS
Vantage SE with fiber optics and Diva software operating
system).
- The FACS Calibur and FACSort have two lasers and can analyze cells based
on 6 separate parameters (side and forward light scatter, and detectors for
4 different fluorescent colors).
- The FACS Vantage SE is a high-speed cell sorter. It has two lasers and
can analyze and sort based on 8 separate parameters (side and forward
scatter and detectors for 6 different fluorescent colors). The FACS Vantage
can simultaneously sort up to four populations of cells. It is also possible
to sort one or more cell types aseptically into 6, 24, and 96 well tissue
culture plates.
- There is a central data analysis software program (FCS Express) on the
departmental network that can be accessed by signing up for its use.
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 Director
William C. Davis
509-335-6051
davisw@vetmed.wsu.edu |
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Protocols and Services Available
- Immunophenotyping of peripheral blood cells
and cell suspensions from tissues from a wide
range of species. Flow cytometric phenotyping is
used diagnostically to determine the cell type
of malignancies, therapeutically to monitor cell
populations, and in research to monitor cell
populations following infection, disease course
and immunization studies. Panels of monoclonal
antibodies for use in specific species are
available from the WSU Monoclonal Antibody
Service Center.
Isolation of PBMC from peripheral blood
Labeling cells with monoclonal antibodies (mAbs)
for Flow Cytometry
- Monitoring of harmful bacteria and protozoan
species in water supplies.
- Isolation of a single cell type from a mixed cell population.
Applications include production of specific T cell clones, enrichment of
cells expressing a specific product such as a monoclonal antibody or
GFP-tagged recombinant protein, and enriching the frequency of a rare cell
type in a population.
- Determination of ploidy of cells using a DNA intercalating fluorescent
marker. Applications include cell cycle analysis and detection of triploid
or tetraploid cells.
- Monitoring of apoptosis.
- Detection of infection by intracellular pathogens.
- Cell viability. Use
of propidium iodide to label dead cells
- Detection of cytokine or interleukin production by cells.
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Detection of intracellular antigens
Intracellular detection of Foxp3
Monoclonal
Antibodies
Panels of monoclonal
antibodies are available for a variety of species. Click on the image of the
species of interest for more information. Antibodies can be purchased from the
WSU Monoclonal Antibody Center.
Technical Support
Technical support is
available to users of this facility. The FACS Calibur and FACSort are
user-friendly flow cytometers that can be used by students, staff, and faculty
without technical assistance after instruction on operation. Technical and
operations support will be provided to researchers who wish to use the flow
cytometers. Training will be provided to researchers who will require more than
occasional use of these instruments. The FACS Vantage requires technical
assistance at all times. Arrangements to schedule use of the cell sorter can be
made by contacting Dr. Davis. Training in the use of the sorter will be provided
to a designated technician or graduate student in laboratories requiring
long-term heavy use of the sorter.
Authorized individuals have access to the core facility at all times during
the week and weekends. Frequent users can obtain key badges. Also, a key to the
room is available in Dr. Davis' laboratory for occasional users of the facility
(Bustad Hall Room 324). Users may reserve the time they will be using the flow
cytometers in advance. Call Dr. Davis' laboratory, 335-6058, to reserve time on
the flow cytometers.
Hourly fees are charged for use of the cytometers. Contact Dr. Davis to
inquire about fees.
Flow Cytometry Intensive Short Course
Duration: 2 weeks on site
Topics covered
1. Preparation of cells for labeling
2. Instruction on how to set up 1, 2, 3, and 4 color flow cytometry panels
3. Protocols for surface and intracellular labeling
4. Instruction on setting up an analytical flow cytometer, including
compensation and collection of multiple colors
5. Data analysis using FCS Express
6. Instruction on setting up and using the FACS Vantage cell sorter
7. Starter set of monoclonal antibodies appropriate to the species of
interest
Contact Information
WSU Monoclonal Antibody Center
Dr. William C. Davis
Veterinary Microbiology and Pathology
PO Box 647040
Washington State University
Pullman WA 99164-7040
Telephone: 509-335-6051
Email:
davisw@vetmed.wsu.edu
Other sites of interest
Other sources include
AbD Serotec and
AbCam.
http://www.vetmed.wsu.edu/tkp/index.aspx The TKP program (taxonomic key
program) is a program designed to assist investigators in the characterization
of new mAbs specific for leukocyte differentiation molecules (LDM) in various
species that are orthologues of LDM fully characterized in humans. Investigators
can use the data base to compare the characteristics of the new mAb-defined
molecule with the characteristics of known molecules, based on molecular weight,
surface orientation, pattern of surface expression on one or more cell types,
and molecular weight. A short description is provided in the data base. There is
a link to a second database that provides information on existing mAbs that
react with the respective CD molecules in one or more species. The antibody
database can be accessed directly through the TKP for investigators attempting
to find mAbs to the CD of interest.
