Flow Cytometry and Human Immune Monitoring Shared Resource

The Human Immune Monitoring Facility (HIMF)

The Human Immune Monitoring Facility (HIMF) is a shared resource within the Arizona Cancer Center that provides comprehensive immune profiling, sample preparation, and clinical trial support. Our mission is to provide technical expertise to clinical investigators and research scientists who want to characterize the immune system by offering up-to-date immune monitoring services for clinical and translational studies.

Our services include multispectral flow cytometry, metabolic analysis, blood services such as PBMC isolation and serum/plasma extraction, and a full spectrum of immunological services. We also offer custom assays upon request and will work with interested investigators to help get their projects up and running. Biobanking services are also available upon request. 

Our equipment includes the Cytek Aurora 5 Laser Spectral Flow Cytometer, Seahorse XFe96 Metabolic Analyzer, BioSys Bioreader-6000 ELISpot analyzer, among others. 

We offer training, assisted, unassisted, and technician operated modalities for all our instruments and will meet investigators at their desired level of engagement. All services are available during regular business hours (M-F 8am - 4pm) with 24 hour access to select instruments arriving in the near future. 

Flow Cytometry Shared Resource (FCSR)

Flow cytometry is a powerful tool that can measure functional and structural characteristics of heterogeneous mixtures of cells and particles in suspension. Flow cytometry also enables the sorting, or physical separation, of these cells into different classes based either on physical characteristics of the cells or on stains, or fluorochromes, which can be attached to individual cells. These fluorochromes are attached to the cells via antibodies, DNA, cytokines, specific proteins or other targets that differentiate the various cell subpopulations in terms of physiology, metabolism, morphology, or other characteristics. The ability to distinguish cell subpopulations is limited only by the ability to attach specific fluorochromes to the cells. Data regarding these subpopulations can then be aggregated, displayed and analyzed via histograms or two-dimensional plots.

The FCSR offers instrumentation, knowledge and skills necessary to sort, quantify and analyze cells and particles in fluid suspension through the use of flow cytometry.The FCSR also provides access to expertise in the interpretation of results, as well as consultation regarding project specific applications and methodologies. FCSR also sponsors UA workshops and seminars which provide information regarding new techniques, cutting-edge instrumentation, and developing applications of flow cytometry.

The cell-sorting application of flow cytometry can physically separate cell populations into different collection tubes. Using this application, the FCSR gives researchers the capability to analyze, and then collect up to four different cell populations distinguished by specific fluorescent markers or light scatter, enumerating differences in physiology, metabolism, morphology, or phenotype.

Instrumentation in the FCSR includes a five-laser BD FACSAria III cell sorter, a five-laser BD LSR II analyzer, a three-laser BD FACSCanto II analyzer, and a four laser Imagestream II, imaging cytometer. The FCSR alsoprovides consulting services to help investigators plan new projects, and several software packages typically used for analysis. Training, to enable unassisted, user-only operation, is available for all instruments. Assisted services are available during regular business hours (9:00 a.m. to 5 p.m., M-F), and an online scheduler, iLAB is used to reserve instuments and staff assistance.

The LSR II analyzer, is located in the MRB, across campus from the FCSR, and is available to trained users, 24/7.

Capabilities and Applications

The techniques of flow cytometry and cell sorting can be applied to many disciplines. Methods have been developed to measure the natural fluorescence of phytoplankton, determine the sex of birds, study the characteristics of sperm, and determine the quality of a metropolitan water supply. Traditional uses of flow cytometry include applications in immunology, hematology, oncology, parasitology, and genetics.

Following is a list of some of the measurable parameters of cells and particles using flow cytometry:

• Relative cell size/shape
• Cytoplasmic granularity (organelles)
• Cell surface and cytoplasmic antigens
• Membrane integrity (viability)
• Apoptosis
• DNA content (ploidy) and cell cycle
• Membrane permeability
• Membrane potential (dye/drug uptake)
• Membrane-bound/cytoplasmic calcium 

Leadership

Hours

Location

Human Immune Monitoring Facility (HIMF)

Monday - Friday
8:00am - 4:00pm                

Arizona Cancer Center
1515 N. Campbell Ave. #0913
Tucson, AZ 85724

Flow Cytometry Shared Resource (FCSR)

Monday - Friday
9:00am - 5:00pm                

Arizona Cancer Center
1515 N. Campbell Ave. #0935
Tucson, AZ 85724