Flow Cytometry and Imaging Shared Resource Facility
William M. Grogan, Ph.D.
Resource director
(804) 828-9519
Frances White
Resource manager
(804) 828-9586
Julie Farnsworth
Cytometry specialist
(804) 828-9586
The instrumentation of the Flow Cytometry and Imaging Shared Resource has been regularly upgraded through seven generations of flow cytometers and two generations of confocal microscopes to the four instruments currently in use: a Beckman Coulter EPICS Elite ESP; a Beckman Coulter XL-MCL; a Dako MoFlo high-speed analyzer/sorter; and a Zeiss LSM 510 Meta imaging system.
The Resource provides a wide range of services, from routine fluorescence analysis to interactive custom design of innovative analysis and sorting protocols, addressing the specific needs of individual investigators. The Resource director closely follows the analyses performed by the core facility, insuring quality control and advising facility users on alternative, often novel, approaches to experimental design and data analysis. This shared resource maintains a stable base of instrumentation, expertise and trained personnel and currently provides services and training for multiple users in 59 research groups from 21 departments at VCU. It has had a major impact on cancer research at Massey Cancer Center for 30 years.
The Resource laboratories and offices are located in Sanger Hall on the VCU Medical Center campus, in close proximity to Massey Cancer Center.
Resources and equipment
The Resource provides a full range of custom flow cytometry and confocal imaging services on a fee-for-service basis employing state-of-the-art instrumentation and highly trained, experienced personnel. Users have the option of running their own analyses, leaving their samples for analysis by Resource personnel or staying with the operators while their samples are processed. Consultation is available at all stages in a research project. Available instrumentation includes:
- Beckman CoulterEPICS Elite ESP (Enhanced Sorting Performance) multilaser flow cytometer. The Elite-ESP is a full-functioned research flow cytometer/sorter which can be configured for many different applications.
- Beckman CoulterElite XL-MCLsingle-laser flow cytometer.The XL-MCL is suitable for the majority of common flow cytometry applications, including immunofluorescence and DNA analysis, cell viability studies, and mitochondrial analyses.
- Dako MoFlo high-speed cell sorter.The MoFlo performs three-color analysis and high-speed sorting, with visible and/or UV excitation.
- Zeiss LSM 510Metaconfocal scanning microscope.The Zeiss LSM 510 Meta is a four-laser confocal imaging system used for fluorescence analysis and imaging of cell cultures or solid tissues, complementing flow cytometry.
Services provided
A variety of analyses are available. These include:
Flow cytometry
- Dual-angle light scatter analysis. Capabilities extend to simultaneous analysis of eight or more different populations.
- Immunofluorescence; one-, two-, three- and four-color for simultaneous quantification of up to four different antibodies. Internal antigens can be quantitated by appropriate fixation.
- DNA and cell cycle analysis.
- Receptor binding. In addition, receptor affinity can be determined by varying ligand concentration.
- Internal glutathione, pH and free Ca+2.
- Viability/cytotoxicity.
- Viable mitochondrial staining using rhodamine 123. This is also suitable for cytotoxicity studies.
- Membrane potential using carbocyanine dyes.
- Sorting of viable cells under sterile conditions.
- Real-time analysis, up to 30 minutes.
- Cells ranging in size from platelets to muscle cells can be analyzed.
- Apoptosis can be followed using a variety of dyes.
Imaging cytometry
- Membrane fluidity by FRAP analysis.
- Gap junctions by FRAP analysis.
- Localization and quantitation of cellular fluorescence.
- Immunofluorescence.
- Quantitation of internal free calcium and internal pH.
- Colocalization by Fluorescence Resonance Energy Transfer (FRET) analysis
Other useful applications include measurement of intracellular glutathione, membrane potential, viable mitochondrial staining for toxicity, binding to cell receptors and cellular uptake of drugs or hormones.
Training classes
The Resource manager periodically conducts training sessions in flow cytometry and confocal imaging to authorize facility users to perform their own analyses.
Service access and charges
Contact flowlab@mail2.vcu.edu or call (804) 828-9586 for more information.
