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, Nanostring GeoMx DSP, Biotek Cytation 1, 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


Sara Centuori, PhD Shared Resource Director Research Asst. Professor, Medicine Phone: (520) 626-3249 Email: smb4@email.arizona.edu	Mark Curry Shared Resource Manager Phone: (520) 621-2047 Email: markcurry@email.arizona.edu

Hours of Operation and Location

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

Links and Resources

See us on the Arizona Research Labs Website
See us on the AZ Cancer Center Website
The Flow Cytometry Shared Resource Equipment Kiosk.

Contacts

Name	Role	Phone	Email	Location
Flow Cytometry Shared Resource	General Facility Contact	520-621-2047	Flow@email.arizona.edu	Leon Levy Cancer Center Room 0935
Core Facility Business Office	Billing Inquiries	520-621-4064	CoreBusiness@email.arizona.edu	Keating Room 200
Human Immune Monitoring Facility	Human Immune Monitoring	520-626-7254	pgundy@arizona.edu	Leon Levy Cancer Center Room 0913

Service list

Name	Description	Price
Cell Culturing and Maintenance	Short term maintenance of cell culture utilizing HIMF facilities. This includes cell culturing for assay prep. We can support a variety of tissues and cells. Per Labor Hour	Internal $45.00 /hr Non-Profit/Educational $60.00 /hr For-Profit $75.00 /hr
Complex Immune Assay	General catch all for immunologic services not listed. Please contact the facility using our Consultation Form to discuss your project. Assays can include: T-Cell proliferation Antigen presentation assay Miltenyi MACS magnetic column separation Please contact us directly to discuss how we can support your project. Per Labor Hour	Internal $75.00 /hr Non-Profit/Educational $110.00 /hr For-Profit $150.00 /hr
ELISA	Enzyme linked immunosorbent assay on a 96 well plate. Custom and kit based ELISA available. Per Each Plate	Inquire
Fixation for CyTOF	Preperation of samples for Mass Cytometry. Per Labor Hour	Internal $45.00 /hr Non-Profit/Educational $60.00 /hr For-Profit $75.00 /hr
HIMF - Staff Labor	Per Labor Hour	Internal $35.00 /hr
Nucleic Acid Purification	Purification of Nucleic Acids using a standardized kit. If a user requests a specific kit not used by HIMF, the user is responsible for providing their kit to HIMF. High throughput 96-well purification methods also available. Inquire for pricing. Per Sample	Internal $35.00 each Non-Profit/Educational $47.50 each For-Profit $60.00 each
PBMC Isolation from ≤60ml whole blood	Isolation of PBMCs using Ficoll gradient separation. Custom separation protocols that match your needs can also be used. Per Each	Internal $150.00 each Non-Profit/Educational $225.00 each For-Profit $300.00 each
PBMC Isolation from ≥60ml whole blood	Isolation of PBMCs using Ficoll gradient separation. Custom separation protocols that match your needs can also be used. Per Each	Internal $205.00 each Non-Profit/Educational $307.50 each For-Profit $410.00 each
PBMC Pellet from CPT Blood Collection Tube	PBMC purification, pelleting, and snap freezing of PBMCs from whole blood after RBC lysis. Per Each	Inquire
Plasma or Serum Extraction	Isolation and aliquoting of plasma and/or serum Per Each	Internal $40.00 each Non-Profit/Educational $60.00 each For-Profit $80.00 each
Sample Handling, Processing and Storage	Storage, processing and handling of biological samples in a -20, -80, or -195 degree environment. Per Labor Hour	Internal $40.00 /hr Non-Profit/Educational $55.00 /hr For-Profit $80.00 /hr
Sample Staining	Staining in preparation for Flow Cytometry or other immunologic assays. Reagents such as staining antibodies and buffers are charged separately. Per Each Sample	Internal $105.00 each Non-Profit/Educational $137.50 each For-Profit $170.00 each
Tissue Dissociation - Mechanical	Mechanical dissociation of solid tissue into single cells in preparation for downstream analysis such as flow cytometry. Tissue inputs can include solid tumor as well as a varity of human and murine tissues. Various preservation techniques are available. Please inquire about your specific project by contacting us through the cosultation form under the reservations tab. Per Sample	Internal $110.00 each Non-Profit/Educational $145.00 each For-Profit $180.00 each
White Blood Cell Isolation	Isolation of total white cells using high speed centrifugation. Per Each	Internal $52.00 each Non-Profit/Educational $75.00 each For-Profit $104.00 each

Search available services: View: by category alphabetically

Name	Description	Price
▼ ► Human Immune Monitoring (14)
PBMC Isolation from ≤60ml whole blood	Isolation of PBMCs using Ficoll gradient separation. Custom separation protocols that match your needs can also be used. Per Each	Internal $150.00 each Non-Profit/Educational $225.00 each For-Profit $300.00 each
PBMC Isolation from ≥60ml whole blood	Isolation of PBMCs using Ficoll gradient separation. Custom separation protocols that match your needs can also be used. Per Each	Internal $205.00 each Non-Profit/Educational $307.50 each For-Profit $410.00 each
PBMC Pellet from CPT Blood Collection Tube	PBMC purification, pelleting, and snap freezing of PBMCs from whole blood after RBC lysis. Per Each	Inquire
Plasma or Serum Extraction	Isolation and aliquoting of plasma and/or serum Per Each	Internal $40.00 each Non-Profit/Educational $60.00 each For-Profit $80.00 each
White Blood Cell Isolation	Isolation of total white cells using high speed centrifugation. Per Each	Internal $52.00 each Non-Profit/Educational $75.00 each For-Profit $104.00 each
Sample Handling, Processing and Storage	Storage, processing and handling of biological samples in a -20, -80, or -195 degree environment. Per Labor Hour	Internal $40.00 /hr Non-Profit/Educational $55.00 /hr For-Profit $80.00 /hr
Nucleic Acid Purification	Purification of Nucleic Acids using a standardized kit. If a user requests a specific kit not used by HIMF, the user is responsible for providing their kit to HIMF. High throughput 96-well purification methods also available. Inquire for pricing. Per Sample	Internal $35.00 each Non-Profit/Educational $47.50 each For-Profit $60.00 each
Tissue Dissociation - Mechanical	Mechanical dissociation of solid tissue into single cells in preparation for downstream analysis such as flow cytometry. Tissue inputs can include solid tumor as well as a varity of human and murine tissues. Various preservation techniques are available. Please inquire about your specific project by contacting us through the cosultation form under the reservations tab. Per Sample	Internal $110.00 each Non-Profit/Educational $145.00 each For-Profit $180.00 each
Sample Staining	Staining in preparation for Flow Cytometry or other immunologic assays. Reagents such as staining antibodies and buffers are charged separately. Per Each Sample	Internal $105.00 each Non-Profit/Educational $137.50 each For-Profit $170.00 each
Cell Culturing and Maintenance	Short term maintenance of cell culture utilizing HIMF facilities. This includes cell culturing for assay prep. We can support a variety of tissues and cells. Per Labor Hour	Internal $45.00 /hr Non-Profit/Educational $60.00 /hr For-Profit $75.00 /hr
ELISA	Enzyme linked immunosorbent assay on a 96 well plate. Custom and kit based ELISA available. Per Each Plate	Inquire
Fixation for CyTOF	Preperation of samples for Mass Cytometry. Per Labor Hour	Internal $45.00 /hr Non-Profit/Educational $60.00 /hr For-Profit $75.00 /hr
Complex Immune Assay	General catch all for immunologic services not listed. Please contact the facility using our Consultation Form to discuss your project. Assays can include: T-Cell proliferation Antigen presentation assay Miltenyi MACS magnetic column separation Please contact us directly to discuss how we can support your project. Per Labor Hour	Internal $75.00 /hr Non-Profit/Educational $110.00 /hr For-Profit $150.00 /hr
HIMF - Staff Labor	Per Labor Hour	Internal $35.00 /hr

Available Equipment and Resources

▼ ► Agilent Seahorse XFe96 Metabolic Analyzer (1)

▼ ► Flow Cytometry (7)

▼ ► Nanostring GeoMx (2)