Flow Cytometry

Overview of Services

The Flow Cytometry Core provides investigators with a resource for quantitative studies of cells, extracellular vesicles, and single-cell gene transcriptome analyses. The Core offers sample acquisition and analysis, cell sorting, and consultation for experimental design, interpretation and troubleshooting.

***Radioactive materials are prohibited.

Consultation

All investigators are encouraged to meet with the Flow Cytometry Core technologists to discuss their experiment prior to ordering antibodies and other reagents. This meeting is often useful in that many potential problems can be averted before critical resources are used. The Core can also provide written protocols for staining, fixation and data analysis methods. The Core technologists are available for assistance with sample acquisition, analysis and presentation. Materials available for reference in the Flow Core lab include several textbooks on the basics of flow cytometry and the Current Protocols in Cytometry. In addition, the Core frequently organizes training seminars at different levels of expertise.

Flow Cytometry Applications

• Free of charge guidance in sample preparation and panel design, and affordable prices for instrument training and data analysis.
• Functional assays utilizing fluorescent probes such as quantification of mitochondrial organelle function, calcium influx and apoptosis, and quantification of expressible fluorescent proteins.
• Up to 45+ color cell surface and/or intracellular (cytokine) immunophenotyping.
• Measurements of soluble factors in biological fluids and cell culture supernatants using bead-based multiplex assays.
• RNA based flow cytometry assays to decipher cell population heterogeneity for precise identification of subsets of interest.
• Extracellular vesicle enumeration and characterization.
• Four-way sterile electrostatic cell sorting of preclinical and clinical samples for functional studies.

 Training

• The LRI Flow Cytometry Core offers training programs for Beginning and Advanced Flow Cytometer Operators. These courses cover the hardware and software of our analytical instrumentation. It is the belief of the Flow Core that greater understanding of the instruments and physics employed in experiments will lead to improved data and experimental design. The Flow Core is dedicated to assisting investigators to attain whatever their desired level of expertise.

• The Basic course introduces users to the hardware and software of the Sony ID7000, Sony Sorter MA900, Cytek Aurora, BD Fortessa, BD FACSymphony A5, or Zetaview. It is not meant to immediately provide complete self-sufficiency but rather to embrace the major concepts needed to begin. It is expected that after this training the user will plan and perform their next experiment with the assistance of Core personnel.

• The Advanced course provides methods and strategies to optimize high-level multicolor flow cytometry. Material covered is derived from the top thought-leaders in the field of flow cytometry tempered with personal experience and practicality. Building multicolor assays is a process that requires time, dedication and tenacity to overcome hurdles that can appear when attempting to measure many parameters simultaneously. When the investigator can patiently build an experiment with a series of small steps and optimally characterize the instrument settings for their particular research, truly spectacular results.

What to Bring

Bring something to take notes on. Samples and supplies will vary greatly based on specific research. Core personnel will help define what is needed on a one-on-one basis. The Flow Cytometry Core is located in NE5-215,  the BD FACS Aria Fusion and Sony MA900 is located in the Flow Cytometry Core Sort Lab at NE3-224.

Equipment

Analyzers

• BD LSRFortessa
• BD FACSymphony A5
• Cytek Aurora
• Sony ID7000
• ZetaView Quatt

Cell Sorters

• ThermoFisher Bigfoot
• BD FACS Fusion
• Sony MA900

Single Cell-Omics

• 10x Genomics

Leadership

Kewal Asosingh, PhD, SCYM(ASCP)
Scientific Director for Flow Cytometry
216-444-0891
asosink@ccf.org

Location and hours of operation

Links and Resources

LRI services

Flow Core Policies

FlowJo Policy

Flow Cytometry Core FlowJo Licensing Policy

Disclaimer:

The Flow Cytometry Core uses FlowJo software and makes it available for individual labs. The core does not represent FlowJo, LLC nor does it profit in any way from our licensing arrangement with the company. Just like any other license sharer, the Flow Core pays full price for the license seats used within the Core.

FlowJo Software Licenses

Use of FlowJo software requires a purchased license. The Flow Cytometry Core uses a Licensing Server to distribute licenses.

Contacts