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Flow Cytometry Core

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) provides training and access to state-of-the-art flow cytometry for biomedical investigators at the Columbia University Medical Center. Among the many documented applications, characterization and quantification of cells for surface protein, cytokine, intracellular transcription factor and signalingmolecules 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

HICCC Site

BD Fortessa

BB1710

BD Fortessa

ICRC 333

BD LSRII

BB1710

BD LSRII

ICRC 333

BD FACSCantoII

PS17-503

BD FACS Calibur

ICRC 333

Cell Sorters

CCTI Site

HICCC Site

BD Influx (BB1710) and P&S Influx (PS 17-503)

BB1710 Tel: 212-304-5649; PS 17-503: 212-305-6562

Aria II

HICCC 312 Tel: 212-851-4574

Sample preparation guideline for the analyzers and sorters.

Leica DMI 6000B

Analysis Workstations

Workstations (PS17-503 and ICRC 333) are equipped with Flowjo and Microsoft Office.

Fees

Instrument regular fee ($/hr) Cancer center member ($/hr)
Sorter (Aria and Influx) 65 65
LSRII (with UV) 60 58
Fortessa (without UV) 55 55
CantoII 50 45
Calibur 52 35
Leica Microscope 25 25

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.

Use of Equipment

Email Siu-Hong Ho 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 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.

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 DERC (Diabetes and Endocrinology 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.