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DC CFAR VIRTUAL LAB OPEN HOUSE
MULTIPARAMETRIC FLOW CYTOMETRY
May 22, 2020
What is Multiparametric Flow Cytometry?
“Multiparameter flow cytometry is a powerful analytical and preparative tool (1,2). It enables the rapid measurement of multiple physical and chemical
characteristics of individual cells or particles as they flow past beams of laser light in a focused fluid stream. Using flow cytometry, defined cell types can be
identified within mixed cell populations and studied separately or within the context of functional intercellular interactions. Flow cytometers that provide cell
sorting capabilities can identify specific cell types and physically separate them in bulk or individually (e.g., through indexed cell sorting (3)), into different
groups for further study.”
“In addition to cell analysis, flow cytometry has also extended to the analysis of subcellular organelles and even chromosomes (4), as well as
numerous molecules present in cellular lysates or biological fluids (5). Flow cytometric bead-based assays are well suited for performing multiplex
immunoassays (5). These assays are capable of measuring multiple analytes within small, sometimes very precious, samples such as biological
fluids obtained from multicellular organisms or cultured prokaryotic and eukaryotic cells (6-9).”
What is Multiparametric Flow Cytometry?
https://www.expedeon.com/application-notes/multi-color-flow-cytometry/
https://www.sinobiological.com/category/fcm-facs-secondary-antibody
https://www.sinobiological.com/category/fcm-facs-fluorescence-measurement
https://www.expedeon.com/application-notes/multi-color-flow-cytometry/
What is Multiparametric Flow Cytometry?
How to design an experiment using Multiparametric Flow Cytometry?
The Multiparametric Flow Cytometry Core Laboratory has a customized BD LSRFortessaTM X-20 with 4 lasers:
- Violet 405 nm
- Blue 488 nm
- Yellow Green 561 nm- Red 640 nm
Includes a High-Throughput System (HTS)
and a working station.
Located at GWU Ross Hall Room 611
How to design an experiment using Multiparametric Flow Cytometry?
Capabilities:
- Up to 18 parameters
- High Throughput System (HTS) units
for running 96-well plates- BD FACSDIVA software v8.0.1
How to design an experiment using Multiparametric Flow Cytometry?
Laser
Filter
Fluorophores
How to design an experiment using Multiparametric Flow Cytometry?
How to design an experiment using Multiparametric Flow Cytometry?
Know your markers before selecting fluorochromes
How to design an experiment using Multiparametric Flow Cytometry?
How to design an experiment using Multiparametric Flow Cytometry?
https://www.bdbiosciences.com/en-
us/applications/research-
applications/multicolor-flow-
cytometry/product-selection-tools/spectrum-viewer
https://bitesizebio.com/13696/introduction-to-spectral-overlap-and-compensation-flow-cytometry-protocol/
How to design an experiment using Multiparametric Flow Cytometry?
How to design an experiment using Multiparametric Flow Cytometry?
How to design an experiment using Multiparametric Flow Cytometry?
Multiparametric Flow Cytometry Service
Laboratory Director
Alberto Bosque-Pardos, PhD, MBA202-994-9696
In-site Core Manager
Amanda Macedo, PhD202-994-0444
Administrative Core Manager
Ayo Awoyemi
Services Provided
o Experimental design
o Supervised use
o Training and subsequent unsupervised use
o Flow Jo analysis training
o Online booking google calendar
Multiparametric Flow Cytometry Service
Lab Rules
o Biosafety Level 2 (BSL-2) and BSL-2*
o Personal Protective Equipment: Lab coat, Gloves, (Goggles)
o No open shoes, shorts
o No food
o Core Service Request Form
https://dccfar.gwu.edu/core-services-request-form
DC CFAR Microgrants
• Small, competitively reviewed grants for $2,500 to $5,000 available to full CFAR member investigators:▪ Core Service Microgrants
o Funding to access services offered by DC CFAR Scientific Cores
o Goal to generate preliminary data to strengthen future DC CFAR pilot award or NIH applications
▪ Publication, Poster Printing and Scientific Travel Micrograntso Funding for DC CFAR pilot award and NIH CFAR administrative supplement
recipients to support publication and presentation of study results
• Short applications reviewed on rolling basis with rapid turnaround on funding determination (roughly 2 weeks)
