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 for experimental design, training in the use of the instrumentation, 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 Cytek Aurora CS spectral cell sorter is equipped with five lasers (355nm, 405nm, 488nm, 561nm, and 640nm), three scatter detectors (FSC, SSC, 405 SSC), and 64 fluorescence channels. The spectral sorter captures the entire light spectrum of a fluorochrome, which in turn allows for larger panels (40+) and the simultaneous use of more fluorophores that would otherwise be incompatible. The Cytek Aurora CS is capable of traditional 2-way sorting into 15 ml tubes, 4-way sorting as well as 6-way sorting into 1.5ml and 5 ml tubes, single cell sorting into 96-well plates, and index sorting. The sorter has a temperature-controlled sample loading station for sample injection (fits 5ml and 15ml tubes) and is completed enclosed within a biosafety level 2 (BSL2) containment cabinet. The instrument is equipped with SpectroFlo™ software and users can build their experiment in the Cytek Cloud and import into the Cytek Aurora CS in the core. The instrument can be used for both data acquisition and cell sorting. Users can be trained by the core to use the aurora for data acquisition; however, cell sorting is performed only by core personnel.

The Bio-Rad ZE5 cell analyzer is a 4 laser, 21-fluorescence parameter, highly automated conventional flow cytometer, with a 4-7-7-3 configuration for blue (488nm), yellow-green (561nm), violet (405nm), and red (640nm) lasers, respectively, which includes 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 offers operator-independent programmable sample loading and data collection for up to 48 tubes and 96 well plates. The instrument is equipped with Everest software. Users can be trained by core personnel to use the ZE5 or can request core personnel to run their samples.

The core also provides two dedicated workstations where users can analyze their data using FlowJo or ModFit software at no charge.

Core Capabilities

The FCCS core supports: detection and quantification of up to 64 cell surface and/or intracellular molecules using spectral flow cytometry; 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 up to six defined subpopulations; single or multi-cell sorting into microwell plates or tubes; indexed single-cell sorting into microwell plates or tubes, spectral unmixing for complex multicolor panels with 40+ parameters; and aerosol containment for biosafety level-2 applications.

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