Flow Cytometry Facility
Description: The Flow Cytometry Facility provides resources for analytical and preparative studies of cells using flow cytometry to the FAMU biomedical researchers. This facility offers critically important technology for gaining information about normal versus disease states. The utilization of in vivo (animals and human) or in vitro studies to characterize structures or visualize molecular markers using various image methods is a powerful tool in clinical and biomedical research. The facility will provide resources for image data acquisition and analysis for radioactive, fluorescent, and photographic samples as well as digital imaging and document production. The facility also offers FACS acquisition and analysis for detecting cell surface or intracellular proteins, measuring apoptosis, cell cycle analysis and progression, and cellular physiology that have both biomedical and clinical applications. The recently purchased SONY SH800 cell sorter will build upon the current technologies within this facility by allowing us to sort select cell populations from heterogeneous cell populations for subsequent analysis. Technical assistance for experimental design, interpretation and troubleshooting will be provided for users that wish to learn the full operation of the instrumentation for self-service analysis, or full service.
Specific Services: Services include: a) Operator assisted acquisition and analysis; b) Operator unassisted analysis and acquisition (for individuals who have attended an instructional class); c) Off-line data analysis; d) Cell cycle analysis; e) Calcium flux analysis; f) Receptor Endocytosis; g) Sample staining and analysis; h) Cytometric Bead Arrays; i) Annexin V for apoptosis; j) Multiplexed analysis of signal transduction; k) Phosphorylation target assays; l) Detection of Cell Surface Markers; m) Detection of Intracellular Markers; n) Sorting cell populations of interest o) preparing specimens (i.e. Hematoxylin & Eosin Staining, Single Cell Gel Electrophoresis staining (Comet Assay), fluorescent conjugates, tissue fixing, and immunohistochemistry); p) image processing; and q) data acquisition and analysis for radioactive, fluorescent, and photographic sample.
1) BD Facscalibur 2.0 Flow Cytometer with dual laser (air-cooled argon 488 nm laser and a red 635 nm diode laser) and 4 color detection capabilities (housed in Flow Cytometry Facility);
2) Sony SH800 Cell Sorter with an integrated 4 laser (488nm/405nm/561/638nm excitation wavelength) engine and 6 freeform PMTs with 96 well and 384 well capabilities in a biosafety cabinet
The Flow Cytometry Facility will be operated by Dr. Selina Darling-Reed, faculty coordinator (email@example.com) and Dr. Ramesh B. Badisa, facility manager (firstname.lastname@example.org or 850-561-2896). They will be responsible for conducting individual and group training sessions, organizing workshops and seminars, and provide assistance to requested operators. The facility is open on all working days by appointment from 8 am to 5 pm. In addition to the samples, all users need to bring their own supplies – namely gloves, tips, BD falcon tubes (12 x 75 mm).
An annual fee of $300 for FAMU-based labs is required for use of the Flow Cytometry Facility. Please note that the number of users per lab does not affect the charge.