Flow Cytometry Research Core: Services

Multiparameter cell phenotyping: 2 to 6 color antibody combinations.

Isolation of phenotypic cell subsets: fluorometric cell sorting.

Analysis of cell activation,proliferation and apoptosis : cell cycle/DNA content using PI, 7-AAD, and/or BrdU, Tunel staining, AnnexinV expression; proliferation using CFSE dye; activation using Ca++ flux, surface expression of CD69, internal expression of Ki-67, cdK, bcl-2, or p53.

Analysis and quantitation of internal proteins: cytokines and chemokines; cell signaling regulatory proteins; HIV regulatory and structural proteins.

Quantitation of molecular promoters and reporter genes: expression of GFP, beta-gal, HSA, murine Thy-1.

Analysis of cells or tissue on slide: multiparameter cell phenotyping and DNA analysis.

Large volume, high-speed cell sorting; and single-cell deposition and cloning.

Live cell sorting of biohazardous samples.

Instrumentation

BD Biosciences FACSCanto is equipped with two lasers: solid state 488 nm and 633 HeNe. It is a fully digital instrument that is capable of 6 color analysis, with enhanced separation and detection of dim fluorescence peaks.

DakoCytomation MoFlo is equipped with three lasers: Coherent Enterprise II, containing both 488 Argon and UV, and a separate Melles Griot 633 HeNe. It is a jet-in-air, high-speed cell sorter capable of event rates in excess of 25,000 e/sec, and up to 9 color combinations. It is located in a BSL-2/3 environment.

Beckman Coulter Epics Elite is equipped with two lasers: 488 Argon and 633 Helium Neon (HeNe). It is capable of 4 color analysis.

BD Biosciences FACStar Plus is equipped with one laser: Coherent Nova 70 488 Argon. It is a low speed cell sorter capable of an event rate of ~7000 e/sec, and 3 color analysis.

CompuCyte Laser Scanning Cytometer is equipped with 2 lasers: 488 Argon and 633 HeNe. This microscope-based instrument serves as a complement to traditional flow cytometers; it provides analysis of cells attached to slides and thin section tissue samples. In addition, it has a Kodak Digital Science camera for producing photomicrographs.