Flow Cytometry Resource Facility

Welcome!

The Flow Cytometry Resource Facility provides support for faculty and student research at Princeton University, as well as for investigators outside of the university at other academic institutions or private companies. In addition to providing access to the flow cytometers and cell sorting services, we provide technical support to assist in experimental design, data analysis/interpretation and presentation as well as in the development of novel flow cytometric and FACS based techniques.

 

Flow cytometry is a laser based analytical tool in which cells or small organisms in suspension flow at high speed past one or more focused laser light sources. Fluorescently labeled probes and dyes can be added to the sample to determine specific physical and/or biochemical properties of the cells. Optical filters and detectors direct and measure the scattered laser light as well as the fluorescence that is emitted from the cell. Various software platforms are used to acquire and analyze the data, which can be exported in several formats for presentation.

 

The facility analyzes and sorts a wide range of cell types and species including human, murine, porcine and insect cell lines as well as primary murine leukocytes and stem cells, bacteria, yeast, C. elegans, and aquatic organisms, which reflects the diversity of the research at Princeton University.

 

Facility Use

The Flow Cytometry Resource Facility provides flow cytometry (FC) and fluorescence activated cell sorting (FACS) services, training for independent use of certain instrumentation, and expert consultation to assist with experimental design and troubleshooting, as well as data analysis/interpretation and presentation.

 

To request training, consultation, cell sorting and/or assisted services, please register with iLab.

 

 

Getting Started with iLab:

• Princeton Clients:  If you already have an account, log in (upper right corner) and go to the Request Services or Schedule Equipment tabs.  If you are a new user, log in (upper right corner) using a valid Princeton ID.
• Non-Princeton Clients:  Please email cdecoste@princeton.edu directly with any questions about use of this facility.  If you already have an account, log in (upper right corner) with approved iLab credentials.  If you are a new external user, Register to setup an iLab account.
• Visit the iLab Help Site or Customer Help Manual for further guidance using iLab's Core Facility Services.

 

About the Facility

The Flow Cytometry Resource Facility is available 24/7 for use of the flow cytometry benchtop analyzers and self-service FACS sorters by fully trained and authorized clients.

Staff assisted services are available Monday - Friday from 10am - 6pm, with after hours sort experiments available by special request.

 

Instrumentation Available

 

Cell Sorters:

• BD Biosciences FACSAria Fusion
• Sony Biotechnology MA-900
• BioRad Laboratories S3e
• Union Biometrica COPAS BioSort

Flow Cytometers:

• BD Biosciences 5-laser SORP FACSymphony A3 w/ High-throughput sampler
• BD Biosciences 5-laser SORP LSRII w/ High-throughput sampler

Particle/Cell Counters:

• Beckman Coulter Z2
• BioRad Laboratories TC-20

CytoSpin Centrifuge:

• ThermoScientific CytoSpin4

 

Commonly Used Applications and Fluorochromes

• Cell phenotyping with fluorescently tagged monoclonal antibodies. Common fluorochromes include: FITC; Alexa 488, 635 and others; PE; PE-Cy5; PE-Cy7; PE-TxRed; PerCP-Cy5.5; PE-Cy7; APC; APC-Cy7; Pacific Blue; AmCyan; BV421; V450; V500, etc.
• Cell cycle analysis and DNA content determination; proliferation; apoptosis/cell death; and cell counting assays: Propidium iodide (PI); Hoechst 33342; Sytox Green; Syto13; Pyronin Y; Acridine Orange; CFDA-SE and others.
• Detection of one or more fluorescent proteins: EGFP, YFP, CyanFP, DsRed, HcRed, mRFP, mCherry, TagBFP, iRFP and many others. The new multi-laser systems make it possible to simultaneously detect four or more fluorescent proteins.
• Functional and biochemical experiments such as calcium flux, glutathione levels, phosphorylation states; measurement of intracellular proteins and transcription factors; bead array assays for cytokines and other secreted compounds; gain of function and loss of function studies and much, much more!

Acknowledgements

We appreciate that you have chosen our facility to conduct your study and would like to remind you to please add a statement along these lines to acknowledge the Flow Cytometry Resource Facility in your publications and presentations:

 

1.  "We thank Christina DeCoste (and/or Katherine Rittenbach, Gabriel Palmieri) and the Molecular Biology Flow Cytometry Resource Facility which is partially supported by the Rutgers Cancer Institute of New Jersey NCI-CCSG P30CA072720-5921."

2.  If you publish using data that was acquired on the BD Biosciences FACSymphony A3 Flow Cytometer, please also acknowledge the funding award used to purchase the system: S10 award 1S10OD028592-01A1 

Leadership

Christina J. DeCoste, Director

Phone: 609-258-1695
Email: cdecoste@princeton.edu

Gabriel Palmieri, Research Specialist

Phone: 609-258-2473
Email: gabrielpalmieri@princeton.edu

 

Location and hours of operation

Hours
Assisted Services: (After hours sorting appointments are available upon request)        Unassisted/Self-Use for approved, trained users:	10am - 6pm                             24/7
Location
Moffett 122 Princeton University, Dept. of Molecular Biology
Princeton, NJ 08544

 

Educational Resources and Webinars

Introductory Flow Cytometry Materials:

5 short videos at:  https://www.thermofisher.com/us/en/home/support/tutorials.html

Other basic videos:  https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-analysis-learning-center/molecular-probes-school-of-fluorescence/flow-cytometry-basics.html

BD Biosciences e-Learning:  https://www.bdbiosciences.com/en-us/support/training/e-learning-courses

• Recommended courses:
  • Introduction to Flow Cytometry (including how cell sorting works)
  • BD FACSDiva™ Software Basics Training

Other BD Biosciences resources

• "Scientist to Scientist" Panel Design Education Videos at https://www.bdbiosciences.com/en-us/support/reagent_education
• Educational videos on the YouTube channel at https://www.youtube.com/channel/UCLeqmZod8vKvrzsK4BS69nA

 Bio-Rad’s “Flow Cytometry Basics Guide” – download here:  https://info.bio-rad.com/Flow-Cytometry-Guides-LP.html?_ga=2.147319159.1924947811.1585686584-1287800828.1585686584

 

Multi-color Flow Cytometry:

BD Biosciences Horizon tour resources – click “Horizon” and “Video Library”:  https://www.bdbiosciences.com/en-us/applications/research-applications/multicolor-flow-cytometry

 

Recently recorded live webinars

BD Harmony Seminar: ‘A Solid Foundation for Simpler and Smarter Panel Design’

• Speaker: Bob Balderas, VP Biological Sciences, BD Biosciences
• Watch the webinar here

Thermo Fisher NY/NJ Webinar Series #1: 'Cell Biology Research at Top Speed: Optimal Tools and Techniques for Flow Cytometry'

• Speaker: Leticia Montoya, PhD
• Watch the webinar here

Thermo Fisher NY/NJ Webinar Series #2: 'RNA Expression Analysis by Flow Cytometry'

• Speaker: Dariusz Stepniak, PhD
• Watch the webinar here

 Memorial Sloan Kettering Cancer Center (Flow Cytometry Core Facility)

• FCS Express and FlowJo special topics and tutorials
• View the previous live webinars here

FlowTex Webinar Series (partnering with MSKCC FCCF)

• View the previous live webinars here

Beckman Coulter Life Sciences

• 'Measuring Single EVs and their Cargo: Sensitive and Specific Vesicle Flow Cytometry'
• Speaker: John Nolan, PhD
• Watch the webinar here

Thermo Fisher NY/NJ Webinar Series #3: 'Novel Flow Cytometry Techniques to Functionally Characterize Immunotherapies'

• Speaker: Chris Lansdorf
• Watch the webinar here

 Current Protocols (Wiley): 'Providing a Solid Foundation for Simpler & Smarter Flow Cytometry Panel Design'

• Speaker: Mirko Corselli, Ph.D, BD Biosciences
• Watch the webinar here

ThermoFisher: 'Novel Fluorescent Assays and Labeling Strategies for Flow Cytometry'

• Speaker: Jolene A. Bradford, MLS, SCYM(ASCP)
• Watch the webinar here

International Union of Immunological Societies: 'Adaptive Immunity: Memory, Protection and Immunopathology in COVID-19'

• Speaker: Andreas Radbruch, Ph.D.
• Watch the webinar here

 Science Magazine: 'Monitoring the immune system to fight COVID-19: CD4 status, lymphophenia, and infectivity'

• Speakers: Andrea Cossarizza, M.D., Ph.D., Maurice O’Gorman, Ph.D., M.B.A., (D)ABMLI, and Lishomwa (Lish) Ndhlovu, M.D., Ph.D. 
• Watch the webinar here

Cytek Bio: 'Applying Panel Design Principles to Investigate COVID-19 Immune Response with the Cytek Aurora'

• Speaker: Dr. Maria Jaimes
• Watch the webinar here

BD Biosciences: 'Chronic Lymphocytic Leukemia Phenotyping with Single-Cell Multiomic Solutions'

• Speaker: Aaron Tyznik, PhD
• Watch the webinar here

The Francis Crick Institute and MSKCC: 'Flow Cytometry: Experimental Set-up in Diva'

• Speakers: Kathy Daniels, Derek Davies, Rui Gardner
• Watch the webinar here
• Princeton LSRII users- begin watching at the 33 minute mark

BD Biosciences: 'Ask the Expert: Learnings and Advancements in High Parameter Flow Cytometry'

• Speaker: Bob Balderas
• Watch the webinar here

ISAC & Cyto University: 'Discovering Immune Features During COVID-19 By Advanced Flow Cytometry'

• Speaker: Andrea Cossarizza, Ph.D.
• Watch the webinar here
• A free Cyto U account must first be created

BD Biosciences: 'Ask the Expert: Multiparameter Flow Cytometry in Measurable Residual Disease'

• Speaker: Michelle McNamara
• Watch the webinar here

Union Biometrica webinars

• To learn about potential applications of our facility's COPAS Biosort large particle sorter, view the webinars at https://www.unionbio.com/

Advanced topics available for free at this time from CYTO U, ISAC's education portal

“On-demand webinars”:  http://cytou.org/store/provider/custompage.php?pageid=7

• “Data Analysis Rigor and Reproducibility Parts 1, 2 and 3”
• “A Guide to choosing fluorescent protein combinations for flow cytometric analysis based on spectral overlap”
• “Step-by-step multi parameter panel design”
• “Cytometry Part A Spotlight: 21-color flow cytometry for immunophenotyping human blood”
• “Cytometry part A spotlight—16 color panel to measure inhibitory receptor signatures from multiple human immune cell subsets” (higher level panel design and analysis)
• “DAFI Directed Automated Filtering and Identification of Cell Populations from PolyChromatic Flow Cytometry Data” (high dimensional cluster analysis intro)
• “Bridging flow cytometry with new technologies” (CRISPR, IPSC phenotyping, RNA Single Cell RNA Seq and Single Cell manipulation prior to RNASeq – useful, but out of date (2014))
• “RNA Flow cytometry” (overview of PrimeFlow (eBio) and SmartFlare (Millipore) products)
• “Integrating flow cytometry and single cell transcriptomics”

Also check out “CYTO Conference Archives” for interesting topics recorded at our annual professional society meeting:  http://cytou.org/store/provider/custompage.php?pageid=10

Panel Building & Antibody Search Tool

 A very useful free service for academic labs that provides antibody search, spectra viewer and panel design tools:  https://fluorofinder.com/

• For panel design:  Select “Design Panel”, Enter ‘Princeton University’ as Institution name, Select “LSRII” or “FACSAria Fusion” to interact with our exact cytometer configurations.
• You can email Tina your designed panel for review directly through this platform.
• Training on utilizing the panel builder is also available through the company via a 15-min web conference.

Recently Recorded webinar: FluoroFinder

• 5 Most Useful Flow Cytometry Panel Design Features on FluoroFinder
• View the webinar here

Live Webinar: Hosted by FluoroFinder

• 5 Most Useful Flow Cytometry Panel Design Features on FluoroFinder
• Tuesday April 28 @ 10 AM EST
• Registration: Click Here

FCS Express recorded webinars

• All previous live webinars have been recorded
• Watch them here

FlowJo University

This site provides tutorials and webinars on analyzing Flow Cytometry data and using FlowJo analysis software.

•  Visit https://www.flowjo.com/learn/flowjo-university/flowjo

Software

FCS Express

• The Flow Cytometry Resource Facility holds a site license for FCS Express
• All Princeton members can use FCS Express Flow Cytometry analysis software, free of charge.
• To gain access, send an email to Gabriel and Tina, or follow the instructions at https://princeton.corefacilities.org/sc/3967/flow-cytometry-resource-facility/?tab=equipment
• Gabriel and Tina are happy to answer any questions you may have as you learn to use FCS Express and perform your Flow Cytometry analysis

 

FlowJo

• FlowJo is offering a free 30-day serial number license to anyone interested.
• First, download the software appropriate for your computer at https://www.flowjo.com/solutions/flowjo/downloads
• Next open FlowJo and copy your computer's "Hardware Address"
• Request a serial number license at https://www.flowjo.com/covid19-license
• You will be sent an email within 10 minutes that has your assigned serial number license
• Open FlowJo. Acept the terms and conditions. Enter your serial number license next to "Hardware Adress". Be patient, it will take a minute to process.
• Press Done, and you can begin using FlowJo.
• Gabriel and Tina are happy to answer any questions you may have as you learn to use FlowJo and perform your Flow Cytometry analysis.

Contacts

Name	Role	Phone	Email	Location
Christina J. DeCoste	Director	609-258-1695	cdecoste@princeton.edu	Moffett 122
Gabriel Palmieri	Research Specialist	609-258-2473	gabrielpalmieri@princeton.edu	Moffett 122

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