Flow Cytometry

  • Remi Creusot, PhD
    • Director
  • Caisheng Lu, PhD
    • Technical Director

Flow cytometry has become the primary tool for the identification of cell populations according to specific parameters, and is therefore employed by an ever-growing number of biomedical scientists. The ability to design, perform and analyze data from multi-parametric flow cytometric experiments requires technical expertise but also immunological expertise to appropriately design the experiment.

The Flow Cytometry Shared Resource, operated jointly by the Herbert Irving Comprehensive Cancer Center and the Columbia Center for Translational Immunology (CCTI), provides training and access to state-of-the-art flow cytometry for biomedical investigators at the Columbia University Irving Medical Center. Among the many documented applications, characterization, and quantification of cells for surface protein, cytokine, intracellular transcription factor, and signaling molecules are the most common experiments performed in the flow cytometry core facility. The Flow core operates at two sites in the Medical Center, on the 17th floor of the Black Building/Physicians & Surgeons (P&S) Building and the third floor of the ICRC building.

Please contact the facility managers for any issues regarding training, assistance, and access with flow including protocol design, instrument operation, data analysis, and troubleshooting. Additional information may be found at the CCTI Flow Cytometry website.

Visit the CCTI Flow Cytometry Website


Instrumentation

Analyzers

BD Fortessa (P&S)

Features: 405, 488, and 633nm lasers, 14 colors
Location: PS17-514

BD Fortessa (ICRC)

Features: 405, 488, 561, and 633nm, 16 colors
Location: ICRC 325

BD LSRII (P&S)

Features: UV, 405, 488, 532, 594, and 633nm lasers, 18 colors
Location: PS17-514

BD LSRII (ICRC)

Features: UV, 405, 488, and 633nm, 18 colors
Location: Outside ICRC-326

BD FACSCanto II (P&S)

Features: 405, 488, and 633nm lasers, 8 colors
Location: PS17-503

Cytek Aurora 5L (P&S)

Features: 355, 405, 488, 561 and 640nm lasers, 40 unique fluorochrome signatures, 12 fluorescent protein signatures
Location: PS17-514

Cytek Aurora 5L (ICRC)

Features: 355, 405, 488, 561 and 640nm lasers, 40+ unique fluorochrome signatures, enhanced small particle detector 
Location: ICRC-325 

Cytek Aurora 3L (P&S)

Features: 405, 488 and 640nm lasers, 30+ unique fluorochrome signatures,  
Location: PS17-503 

Cell Sorters

BD Influx (P&S)

Features: 405, 488, 561, and 633 lasers, 15 colors
Location: PS17-514

BD Influx (P&S)

Features: 405, 488, 561, and 633 lasers, 15 colors
Location: PS17-503

BD Influx (ICRC)

Features: 405, 488, 561, and 633 lasers, 15 colors
Location: ICRC 301

BD Aria II (ICRC)

Features: 405, 488, 561, and 633 nm, 14 colors
Location: ICRC 301

Sony MA900 (ICRC)

Features: 405, 488, 561, and 638 nm, 12 colors
Available 24/7 after completion of training and demonstration of proficiency
Location: ICRC-326

Analysis Workstations

Workstations in PS17-503 and ICRC 320A with FCS Express, Flowjo, and Microsoft Office installed are available to all the core users for free. Please sign up through the iLab.

Flow Analysis Data Packages

FCS Express: Please contact Caisheng Lu (cl3055@cumc.columbia.edu) to set up a license.


Acknowledgements

As the recipient of four NIH awards, we are required to acknowledge the NIH funding source in all publications (manuscripts, abstracts, presentations, etc.). Please use the below wording in every publication which results from the use of the flow core instruments.

  • If you use the 5L Aurora: 
    • "Research reported in this publication was performed in the CCTI Flow Cytometry Core, supported in part by the Office of the Director, National Institutes of Health under awards S10OD030282. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health."
  • If you use our Influx: 
    • "Research reported in this publication was performed in the CCTI Flow Cytometry Core, supported in part by the Office of the Director, National Institutes of Health under awards S10OD020056. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health."
  • If you are a Cancer Center member and/or use equipment at the ICRC site: 
    • "These studies used the resources of the Herbert Irving Comprehensive Cancer Center Flow Cytometry Shared Resources funded in part through Center Grant P30CA013696."
  • If you are a DRC (Diabetes Research Center) member: 
    • "These studies used the resources of the Diabetes Research Center Flow Core Facility funded in part through Center Grant 5P30DK063608."

Investigators who do not comply with this NIH requirement may lose access to the core services. Please email us the Pubmed ID for the publication with the above acknowledgement.


Locations and Contacts

The Flow Cytometry facility has two sites:

CCTI site

P&S Building 17-514
650 W. 168th Street
Tel: (212) 304-5649

P&S Building 17-503
630 W. 168th Street
Tel: (212) 305-6562

HICCC Site

Irving Cancer Research Center
1130 St. Nicholas Avenue, Rooms 301, 325, 326, 320A
Tel: (212) 851-4950

Contacts

Caisheng Lu, MD, PhD
Technical Director
Room BB1706F (office)
Tel: 212-605-6596
Email: cl3055@cumc.columbia.edu

Ching-Yuan Chen, PhD
Associate Manager
Room ICRC-320A (office) and ICRC-301 (sorter)
Tel: (212) 851-4576
Email: cc3538@cumc.columbia.edu

Wan-I Kuo, MS 
Staff associate 
Room PS17-503 (sorter)

Tel: 212-305-6562 
Email: wk2355@cumc.columbia.edu


User Fees and Policies

Use of Equipment

Email Caisheng Lu to discuss which instrument best fits the experiment and training will be arranged accordingly. If the user already has experience in operation of the instrument, a 5-10 minutes briefing sessions about the general maintenance of the analyzers would be given. Cell sorter generally is operated by dedicated core facility staff only, but training is available for Aria-II self-operation. Please contact the core for generating the online instrument reservation account.

Reservations

For those who affiliate with the CCTI, Department of Medicine, DRC, SDRC, you can reserve our instrument 3 weeks in advance. Users from other departments can only reserve 48 hours in advance. All reservations are made through iLab. We define your affiliation based on the following:

  • CCTI faculty
  • Department of Medicine research faculty 
  • DRC research interest groups
  • Department of Dermatology research faculty
  • Cancer Center

If you don't find yourself among researchers listed in the above web pages, but you belong to one of the groups, please contact the core manager.

Data Policy

The flow core DOES NOT backup any data, and to facilitate the smooth operation of the Diva and other flow software, deletes all .fcs files and templates frequently WITHOUT prior notice. It is the SOLE responsibility of all flow core users to backup their data RIGHT AFTER the data acquisition. All users must temporarily save their data to the corresponding network drive (non-CCTI or HICCC data server). The network drive is for data transfer ONLY, not for storage (even 1 day), the user must back up the data IMMEDIATELY with their own device. The data in network drive are erased every few days, again, WITHOUT prior notice. Further instructions for the network drive can be found here.

Getting Started

  • Web-based theory session (2 hours) covers the principle of flow cytometry, components of flow cytometry (optics, fluidic, electronic), optical measurement, data analysis, and sorting.
  • Consultation session will be conducted on panel design and protocol suggestion.
  • Fluorofinder panel design web-based tool for CCTI/HICCC cytometers and instruction.
  • The hands-on session (2 hours) includes the instrument startup/shutdown, optical filter selection, software usage, PMT voltage adjustment, data storage, and troubleshooting.

User Fees

 

Instrument Regular Fee ($/hr) Cancer Center Member ($/hr)
Sorter (Aria and Influx) (staff-operated) 80 80
Aria-II (self-operated) 68 68
Sony MA900 (self-operated) 70 70
LSRII (with UV) 60 58
Fortessa (without UV) 55 55
CantoII 50 45
Leica Microscope 25 25
Aurora (5 lasers) 75 75
Aurora (3 lasers) 60 60


The minimum charge for the analyzers is 30 minutes. The reservation time and actual usage are compared and longer duration would be charged. A fee of 100% is charged when the cancellation occurs less than 2 hours of the scheduled time. For Influx cell sorter, the minimum reservation is 1 hour. A fee of 100% is charged when the cancellation occurs less than 72 business hours beforehand.


Online Reservations

The Flow Cytometry Shared Resource uses the iLab Core Management System for service requests and billing. Before you can request services, you must log into iLab and be approved as a lab member.