Fluorescence-Activated Cell Sorting/Flow Cytometry Shared Resource LaboratoryFACS/FCMSRLCore B Smooth Muscle Plasticity COBREDirector: Doug Redelman, Ph.D.
Services • Assistance for Flow Cytometry experiments • Designing relevant protocols • Performing the experiment, i.e., labeling cells, etc. • Analyzing results • Testing antibodies • Isolating cells by FACS • Assistance in grant preparation
Applications • Analytical flow cytometry (FCM) • Cell identification by immunofluorescence both to surface and intracellular components • DNA content & cell cycle analysis • Cell proliferation, e.g., CFSE dilution • Cell viability &/or apoptosis measurements • FACS • Population sorting (antibody &/or Tg cells) • Sorting single cells
FCM/FACS Instruments • Analytical Flow Cytometer – BDBiosciences LSR II • 4 Lasers - 405nm, 488nm, 561nm & 640nm • 18 Fluorescence Detectors • High Throughput Sampler (HTS) – plate loader • FACSFlow fluid management • Cell Sorter – BDBiosciences FACSAria II • 3 Lasers - 355nm, 488nm & 640nm • 11 fluorescene detectors • 4 way sorting of populations • ACDU - sorting into plates
Interfacing with Other Labs • Microscopy – Relate single cell properties with tissue morphology • Proteomics – Populations of cells can be sorted for protein analyses as an alternative to Western blots • Genomics • Populations of cells for expression analyses, deep sequencing, etc. • Single cells for analyses in SCME Lab • Pilot projects in COBRE Phase III