Flow Cytometry


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 Core of the Columbia Center for Translational Immunology (CCTI) and Herbert Irving Comprehensive Cancer Center (HICCC) 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.

Please contact the facility staff for any issues regarding training, assistance and access with flow including protocol design, instrument operation, data analysis, and troubleshooting.

Analyzers

CCTI Site

  • BD Fortessa
    • 405, 488 and 633nm lasers, 14 colors
    • High throughput sampler
    • Location: P&S 17-514
  • BD LSRII
    • UV, 405, 488, 532, 594 and 633nm lasers, 18 color
    • Non-default dye filter change suggestions
    • Location: P&S 17-514
  • BD FACSCantoII
  • Cytek 5L Aurora
    • 355, 405, 488, 561 and 640nm lasers
    • 64 channels, 40+ unique fluorochrome signatures, 12 fluorescent protein signatures
    • Operation manual and fluorochrome selection guideline available. Contact Caisheng Lu

    • Location: PS17-514
  • Cytek 3L Aurora
    • 405, 488 and 640nm lasers
    • 38 channels, 30+ unique fluorochrome signatures, 8 fluorescent protein signatures
    • Operation manual and fluorochrome selection guideline available. Contact Caisheng Lu
    • Location: PS17-503

View the Configuration Charts

HICCC Site

  • BD Fortessa
    • 405, 488, 561 and 633nm, 16 colors
    • Location: ICRC 333
  • BD LSRII
    • UV, 405, 488 and 633nm, 18 colors
    • Location: ICRC 333

View the Configuration Charts

Cell Sorters

CCTI Site

View the Configuration Charts

HICCC Site

View the Configuration Charts

View the Sample Preparation Guideline For Analyzers and Sorters

Leica DMI 6000B

Analysis Workstations

Workstations in PS17-503 and ICRC 333 with FCS Express, Flowjo, and Microsoft Office installed are available to all core users for free. Please sign up through iLab.

Flow Data Analysis Packages

FCS Express: Please contact Caisheng Lu to set up a license, training will be provided for free upon request.

Fees

Sorter (Aria and Influx)

Regular Fee ($/hr): 70

Cancer Center Member ($/hr): 70

Diabetes Research Center Member ($/hr): 68

LSRII (with UV)

Regular Fee ($/hr): 60

Cancer Center Member ($/hr): 58

Diabetes Research Center Member ($/hr): 58

Fortessa (without UV)

Regular Fee ($/hr): 55

Cancer Center Member ($/hr): 55

Diabetes Research Center Member ($/hr): 53

CantoII

Regular Fee ($/hr): 50

Cancer Center Member ($/hr): 45

Diabetes Research Center Member ($/hr): 48

Leica Microscope

Regular Fee ($/hr): 25

Cancer Center Member ($/hr): 25

Diabetes Research Center Member ($/hr): 23

5L Aurora

Regular Fee ($/hr): 75

Diabetes Research Center Member ($/hr): 73

3L Aurora

Regular Fee ($/hr): 60

Diabetes Research Center Member ($/hr): 58

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 the cell sorters, the minimum reservation is 1 hour. A fee of 100% is charged when the cancellation occurs less than 72 business hours beforehand. Assisted use of analyzer instruments will be charged at an additional $60 per hour. A charge for reagents provided by the Core will also be applied. For example, Compensation Beads will be charged $1 per sample.

Flow Cytometry Core data on a computer screen

Use of Equipment

Email Caisheng Lu (CCTI Site) 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 is operated by dedicated core facility staff only. Please contact the core for generating the online instrument reservation account.

Reservations

For those who affiliate with the CCTI, Cancer Center, Department of Medicine, DERC, or 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:

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 director.

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. For CCTI members, save all data to your CCTI P: share or OneDrive. Non-CCTI members, use your CUIMC provided OneDrive directory here: https://cumccolumbia-my.sharepoint.com/. The data in network drive are erased every few days, again, WITHOUT prior notice.

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.

Acknowledgement

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 our LSRII

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 S10RR027050. 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

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 Diabetes Research Center member

These studies used the resources of the Diabetes and Endocrinology 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.

Co-Authorship for the Flow Core Staff

Please list our flow core staff members as a co-author if we're involved in one of the following areas (based on the authorship guidelines from the Association of Biomolecular Resources Facilities published in May 2010):

  1. Conception, design of project, critical input, or original ideas;
  2. Acquisition of data, analysis and interpretation, beyond routine practices;
  3. Draft the article or revise it critically for intellectual content;
  4. Write a portion of the paper (not just materials and methods section);
  5. Intellectual contribution;
  6. Final authority for the approval of article.

Please discuss with the your lab members and the flow core staff for their involvement before your manuscript submission.

Our Team