SLIDE 1
DC CFAR VIRTUAL LAB OPEN HOUSE MULTIPARAMETRIC FLOW CYTOMETRY
May 22, 2020
SLIDE 2 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).”
SLIDE 3 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
SLIDE 4 https://www.expedeon.com/application-notes/multi-color-flow-cytometry/
What is Multiparametric Flow Cytometry?
SLIDE 5
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 6 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?
SLIDE 7 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
SLIDE 8
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 9
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 10
Know your markers before selecting fluorochromes
SLIDE 11
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 12
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
SLIDE 13
https://bitesizebio.com/13696/introduction-to-spectral-overlap-and-compensation-flow-cytometry-protocol/
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 14
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 15
How to design an experiment using Multiparametric Flow Cytometry?
SLIDE 16 Multiparametric Flow Cytometry Service
Laboratory Director Alberto Bosque-Pardos, PhD, MBA 202-994-9696 abosque@gwu.edu In-site Core Manager Amanda Macedo, PhD 202-994-0444 amacedo@gwu.edu Administrative Core Manager Ayo Awoyemi
- luwaseun.awoyemi@howard.edu
SLIDE 17 Services Provided
- Experimental design
- Supervised use
- Training and subsequent unsupervised use
- Flow Jo analysis training
- Online booking google calendar
Multiparametric Flow Cytometry Service
Lab Rules
- Biosafety Level 2 (BSL-2) and BSL-2*
- Personal Protective Equipment: Lab coat, Gloves, (Goggles)
- No open shoes, shorts
- No food
- Core Service Request Form
https://dccfar.gwu.edu/core-services-request-form
SLIDE 18 DC CFAR Microgrants
- Small, competitively reviewed grants for $2,500 to $5,000
available to full CFAR member investigators:
▪ Core Service Microgrants
- Funding to access services offered by DC CFAR Scientific Cores
- Goal to generate preliminary data to strengthen future DC CFAR pilot award or NIH
applications
▪ Publication, Poster Printing and Scientific Travel Microgrants
- 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
- n funding determination (roughly 2 weeks)
