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Flow Cytometry Core - Core D


Core Director:
Doris Tse, Ph.D.
Phone: 212-263-8848
Email: Doris Tse, Ph.D.
First time users should email the Core Director who will
assign your project to one of our staff.

Core Staff:

Core Location:
Bellevue Hospital, OB-C&D 6/F

Instruments:
FACSAria
Recently installed 5 laser (UV, violet, blue, YAG, red) cytometer capable of detecting 17 different emissions. Cells are sorted at flow rates of 70,000 events/sec into 4 distinct populations. It is equipped with an aerosol containment unit, an automated cell deposition unit for isolating a preselected number of cells for cloning, limiting dilution studies or microscopy, and a recirculator for temperature sensitive experiments. This instrument is supported by a power-backup system and can be used for the analyses or sorting of viable cells of primate origin, or non-primate cells exposed to microbial agents. It is located in OB-C&D Room 648.

FACSVantage SE
Equipped with an argon ion laser, a mixed-gas Spectrum laser (UV capable), and an air-cooled HeNe laser. As many as 7 different emissions can be detected.  It is also equipped with an automated cell deposition unit and an aerosol containment unit. This instrument is located in a BL3 laboratory in NB16E21, and will be used for the analyses or sorting of samples containing airborne pathogens. It is scheduled to be replaced by the FACSAria pending construction of a BL3 laboratory in Old Bellevue.

FACSCalibur
4-color cytometer for clinical diagnostics, immunophenotyping, lymphocyte counts, and cell cycle analyses. This instrument is available for hands-on use. It is located in OB-C&D Room 644.

Luminex 200
This instrument utilizes xMAP technology to simultaneously assay up to 100 analytes in a single well of a microtiter plate. Polystyrene microspheres are coded using a mixture of red and infrared fluorophores to form 100 distinct sets. Separate bead sets are coated with reagents that allow the capture of specific analytes contained in a sample, which are subsequently quantified using detection reagents conjugated to a different fluorophore, usually PE. Within the Luminex 200 analyzer, lasers excite the dyes that identify each microsphere, and any reporter dye captured by them during the assay. BeadView analysis software provides automated curve fitting capabilities to generate standard curves and calculate assay results for each analyte. This instrument is located in OB-C&D Room 645.

Services and Fees:
Clinical Testing
Our laboratory is NYSDOH and ACTG/IQA certified.

T/B/NK and CD3/CD4/CD8 subset determination: $52.70 per test 
Other available markers include CD5, CD16, CD25, CD45RA, CD69, γδTCR: $60.00 per test

FACS Analyses and Cell Sorting

  • Immunophenotyping leukocytes from human, mice, and nonhuman primates
  • Leukocyte counts for human isolates
  • Immunophenotyping highly autofluorescent cells, such as tissue macrophages, fibroblasts and osteoclasts.
  • Detection of intracellular cytokines and nuclear antigens
  • Detection of cell proliferation and apoptosis
  • Detection of reactive oxgen species
  • Measurement of Ca++ mobilization
  • Detection of selectable phenotypes on transfected cells
  • Use of FRET to monitor changes in the proximity of molecules on the cell surface, or to detect cell fusion.
Rate on FACSCalibur: $75.00 per hour
Rate on FACSCalibur, hands-on: $50.00 per hour
Rate on FACSAria or FACSVantage: $100.00 per hour
Sterilization for Cell Sorting: $100.00
Biohazard Decontamination after Analyses or Sorting: $100.00
Cell Processing and Labeling: $25.00 per hour plus cost of reagents

Multiplex Analyte (MPA) Detection
Measurement of multiple user-selected analytes in a single 60 µL specimen of serum, plasma, lavage fluid, urine, or culture supernatant. We currently offer analysis of GM-CSF, IFN-α, IFN-γ, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p40, IL-13, IL-15, IL-17, IP-10, MDC, Mig, TARC, TGF-β, TNF-α, sFasL, and VEGF. The core will consider other commercially available analyte detection systems on request.

Cost per specimen (human analytes only) assayed in duplicate: $20.00 for 2 plexes plus
$4.50 for each additional plex.
Minimum fee: $300.00 per assay.
Rate on Luminex, hands-on: $100.00 per 96-well plate


BD Biosciences Contract

Orders placed through our core will receive a 35% discount for BDIS reagents and a 20% discount for BD Pharmingen reagents, independent of whether such reagents are used for flow cytometry.