Flow Cytometry (FCCS)

Overview of Services

Introduction

The Flow Cytometry and Cell Sorting (FCCS) Core was created to provide the UTHSC and Memphis research community with access to state-of-the-art instruments, expertise, instruction, and assistance with experimental design and data analysis for multicolor flow cytometry and cell sorting, including indexed single-cell sorting. Services include one-on-one consultation with internal investigators at no charge for experimental design, training in the use of the instrumentation (hourly rate), and software resources. The Core Director and the Flow Cytometry Specialist, both highly experienced immunologists and flow cytometry and cell sorting experts, are also available to analyze investigators’ data (hourly rate). Together, these activities are part of the service, educational, and academic missions of the FCCS core.

Equipment and Software

The BD Biosciences FACSAria II cell sorter is equipped with four lasers and 12 fluorescence detectors, in addition to forward (FSC) and side (SSC) scatter detectors. The 100 mW, 488 nm blue diode laser has 5 fluorescence, SSC, and FSC detectors. The 30 mW, 638 nm red diode laser has three fluorescence detectors. The 50 mW, 405 nm violet diode laser has two fluorescence detectors, and the 20 mW, 355 nm solid-state UV laser has two fluorescence detectors. The sorter has two-and four-way sort capability into tubes or microtubes. The sorter is also equipped for indexed, single-cell sorting or multiple cell sorting into microwell plates or onto microscope slides. The sorter has temperature controlled sample injection and collection chambers within a biosafety level-2 (BSL2) containment cabinet.

In September 2016, a new high-performance ZE5 (formerly known as the YETI) flow cytometer was purchased to replace the BD Biosciences LSR II. New instrument specifications include a four-laser, 21-fluorescence parameter highly automated flow cytometer, with a 4-7- 7-3 configuration for blue, green, violet, and red lasers, respectively, supporting detection of popular “fruit” dyes and standard FSC and SSC light detection. The instrument also has the capability for small particle detection (exosomes, subcellular particles, and bacteria) and will offer operator-independent programmable sample loading and data collection for up to 48 tubes. For comparison, the LSR II offered three lasers with 9 fluorescence, FSC, and SSC detectors. In addition, the Everest software is more user-friendly than FACSDiva (BD Biosciences).

Core Capabilities

The FCCS core supports: detection and quantification of up to 21 cell surface and/ or intracellular molecules; detection and quantification of GFP- or mCherry-labeled protein expression; measurement of Ca2+ flux during cell signaling among different cell subpopulations; DNA quantification, cell cycle analysis, and apoptosis detection and quantification; detection and quantification of exosomes; high-speed sorting of viable or fixed eukaryotic or prokaryotic cells into four or fewer defined subpopulations; single or multi-cell sorting into microwell plates or onto microscope slides; indexed single-cell sorting into microwell plates or onto microscope slides; and aerosol containment

Leadership

Location and hours of operation

Links and Resources

1. Flow Cytometry and Cell Sorting Laboratory (FCCS) 
2. FCCS Brochure
3. FCCS Core Price List