Flow Cytometry, Microscopy, and Shared Equipment (CCTI)

Flow Cytometry

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. The CCTI Flow Core includes three analytic flow cytometers and two flow sorters.

 

The analytic flow cytometers include the BD FACSCantoII, BD Fortessa, BD LSRII which acquire data up to 10, 16, 21 parameters, respectively. While CantoII and Fortessa meet the needs of many basic experimental needs, our LSRII is among the most advanced cytometers. The LSRII harbors 6 lasers (355, 405, 488, 532, 594 and 633nm), which are able to excite a large fraction of commercially available fluorochromes (colors). In addition to the wide spectrum of lasers, the LSRII is equipped with the 96-well plate reader which allows high throughput analysis of 96 samples in short duration. This high throughput capacity makes projects such as tissue mapping and drug screening possible.

 

The 4-laser BD Influx cell sorter can sort on 17 parameters and provide 4 populations of cells. Importantly, the BD Influx meets BSL 2 biosafety specifications for viable human cell sorting. With the BD influx cell sorter, the populations of interest can be isolated for downstream experiments such as cell expansion/functional studies and DNA/RNA characterization. Please contact the facility staff for any issues regarding training, assistance and access with flow including protocol design, instrument operation, data analysis, and troubleshooting.

 

Contact

Caisheng Lu

Tel: (212) 305-6596
Email: cl3055@cumc.columbia.edu

Wei Wang

Tel: 212-851-4950; 212-851-4603
Email: ww2351@cumc.columbia.edu

Location and hours of operation

Hours Location

Hours Staffed: Monday-Friday 10:00am - 6:00pm

Hours Open: 24/7 

P&S, PS17-514
630 West 168th Street
New York, NY 10032

P&S, PS17-503
630 West 168th Street
New York, NY 10032

 

Links and Resources

  1. CCTI flow core

 

In Vivo Multiphoton/Confocal Microscopy Core

Our in vivo multiphoton/confocal microscopy is advantageous in the following analyses:

  • Video-rate, or 3D time-lapse movie for the analysis of dynamic cellular behavior in live animals;
  • 3D analysis of cell location in live animals;
  • Subcellular-resolution imaging with weak fluorescent signals in deep tissues is made possible by a multi-photon laser and 2 non-descanned detectors; 
  • Multicolor imaging (theoretically over seven colors) is made possible by five single-photon lasers (458, 488, 514, 561, 633nm), one multi-photon laser, and seven detectors; 
  • Fast scanning of a large area with a resonance scanner (over 60 frames per second) is advantageous for the analysis of rare cell populations (e.g. stem cells).

 

Leica TCS SP8 has been designed for confocal microscopy with optimal photon efficiency and high speed. All optical components are matched towards preserving fluorescence photons for image contrast and to improve cell viability in live cell imaging. Backing up this sensitive detection are a high speed scanning system with up to 428 frames per second, large field of view of field number 22 and accelerated Z-stacking by a novel mode for the SuperZ galvanometer called Galvoflow. This suits the instrument for the most demanding samples, highest speed while offering full confocal resolution. Features include:

  • Objectives
    1. 40X NA=1.1 water (#11506342)
    2. 25X NA= 0.95 water (#11506340)
    3. 10X NA= 0.3 (#11506505)
  • Excitation Lasers
    1. Multiphoton laser: 690 – 1040nm (Spectra Physics Mai Tai-DeepSee)
    2. Single photon lasers: 458, 488, 514, 561, 633nm
  • Image Detection: Point scanning with 3 photomultipliers (PMT), 2 hybrid photodetectors (HyD) and 2 Non-descanned detectors (NDD, for 2 photons detection only)
  • Epi-Fluorescence Filters: DAPI, GFP, Cy3
  • XYZ-dimension: Motorized. 

 

 

Contact

Joji Fujisaki
Phone: (212) 304-5695
Email: jf2819@cumc.columbia.edu

 

Location and hours of operation

Hours Location
By appointment only  

Black Bldg, BB828
650 West 168th Street
New York, NY 10032

 

Links and Resources

  1. CCTI in vivo microscopy core

 

Shared Equipment Core

We have the following for CCTI members and collaborators:

  • Cell freezer BB1734
  • Cytospin BB1511
  • Electroporator BB1511
  • Cryostat #1 BB1707
  • Sorvall Highspeed centrifuge BB1707
  • Ultracentrifuge BB1527
  • Elisa reader PS17-503
  • Biotek Plate reader PS17-503
  • Elispot machine PS17-503
  • ABII QOCR PS17-503
  • Animal irradiator RS2000 BB2001

 

 

Contact

Dr. Anette Wu - CCTI operation director
Phone: (212) 304-5695
Email: aw2342@cumc.columbia.edu