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Flow Cytometry Core Facility
Cancer Research Center
Georgia Health Sciences University 

William King 
Manager

 

 

The Center Flow Cytometry Core Facility is a Cancer Center-subsidized shared resource. Its function is to provide high-quality, cost-effective, state-of-the-art flow cytometry and multiparameter cell-sorting instrumentation and associated expertise and services to all investigators on the campus.

The facility was incepted when the Cancer Center building was inaugurated in 2006 and was created to serve the specific needs of Cancer Center members, however, access was eventually opened to all campus researchers because technologies exist in the core that are not otherwise available on the campus and it is otherwise not cost effective or sustainable to duplicate the more expensive and complex equipment that the facility maintains. The laboratory is BSL2 capable, however, it currently functions as a BSL1-level facility. The exception is the sorter flow cytometer, which is housed in a different area, which is BSL2+ capable.

The laboratory is located in The Cancer Research Center, which as the CN mailing code designation, room 4127. Inquiries concerning access to the facility should be directed to the manager, William King.

What is Flow Cytometry?

Flow cytometry is a technology that measures and analyzes the optical properties of mono-dispersed single particles, such as mammalian cells, bacteria, picoplankton, microbeads, yeast, platelets, nuclei and other similarly-sized particles, passing single file through a focused laser beam.

The laser can excite fluorophores that have been used to mark various molecules or physiological functions of the particles. The use of fluorophores with different fluorescence characteristics, multiple lasers and multiple photo detectors allows flow cytometers to measure many characteristics of each particle simultaneously.

An important feature of flow cytometry is that large numbers, for example thousands of particles per second, are analyzed and therefore provide a statistically significant picture of a specimen's physical and biochemical make-up.

To perform these functions, cytometers require a combined system of:

• Fluidics that introduces and restricts the particles for interrogation.
• An excitation source and collection optics to generate and collect thelight signals.
• Electronics that convert the optical signals to proportionate electronic signals and digitize them for computer analysis.

A further strength of flow cytometry, unlike fluorescent microscopy, is that it is able to distinguish between multiple closely emitting fluorophores, such as eGFP and eYFP, in concurrently expressing cells if the appropriate optics and experiment controls exist. If sort criteria have been set up, some flow cytometers can physically sort selected sub-populations.

Optical signals consisting of laser light scatter and fluorescence are sequentially generated by each single particle and consist of the following:

• Low angle forward laser light-scatter (FSC) intensity, approximately proportional to cell diameter
• Side (also known as orthogonal, 90 degree) laser light-scatter (SSC) intensity, approximately proportional to the quantity of granular structures within the cell
• Fluorescence intensities at many wavelengths (FL1, FL2, FL3, etc.)

Common Applications

Common applications of flow cytometry include the analysis and/or sorting of experiments studying:

• Absolute cell counts
• Apoptosis (intracellular homeostasis, free radical production, chromatin condensation, DNA fragmentation, plasma membrane integrity, mitochondrial function, caspase activation, phosphatidylserine translocation, etc.)
• Cellular signal transduction (calcium flux, cytokine studies, intracellular pH and glutathione measurements, cell cycle analysis, cellular transport assays, drug uptake/efflux assays, fluorescent resonance energy transfer [FRET])
• Characterization of multi-drug resistance (MDR) of cancer cells
• Co-localization *
• DNA (cell cycle analysis, cell kinetics, proliferation, etc.)
• Fluorescent in situ hybridization (FISH) *
• Immunophenotyping of nuclear, intracellular and cell surface antigens
• Rare event detection
• RNA
• Single cell cloning
• Spot counting for viral load *, subcellular compartmentalization * and synapse activity *
• Transfection efficiencies
• Translocation *
• Viability

  * signifies applications specific to the ImageStream^X

 

Cancer Center Flow Cytometry Core Facililty Oversight Commitee

• William King
• Bill Dynan
• Lei Huang
• LesleyAnn Hawthorn
• MingQiang Ren

 

William King is soley responsible for the content of this site. Comments, concerns and questions regarding it should

be addressed to him.