1. Fluorescence-Activated Cell Sorting/Flow Cytometry Shared Resource Laboratory FACS/FCMSRL Core B Smooth Muscle Plasticity COBRE Director: Doug Redelman, Ph.D.

2. 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

3. 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

4. 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

5. 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