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 Operator:
Michael Gregory

Research Associate:
Amitav Mohanty, PhD
Xiao Li, PhD

Clinical Assistant:
Joon Kim, MD

Student/Interns:

• Igor Eyzner
• Vlad Tsaltskan
• Alex Ha

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

Instruments:
FACSAria II
Recently installed 5 laser (UV, violet, blue, YAG, red) cytometer capable of detecting 15 different fluoresences. Cells are sorted at flow rates of 50,000 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.

Biotek Synergy HT
A Biotek Synergy HT is a plate reader for detection of colorimetric products from conventional ELISA, as well as fluorimetric compounds. It is used to measure soluble analytes for which xMAP kits are not available.

Biotek ELx50 Strip Washer
An automated plate washer, used for ELISA.

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

• One to eight color immunophenotyping. Human specimens include unfractionated whole blood, bronchoalveolar (BAL) and vaginal lavage fluid, and isolates from tissue biopsies. Murine specimens include cells from spleen, thymus, lymph node, bone marrow, and BAL fluid. Other specimens include an assortment of cell lines from various species. Reagents used include fluorophore-conjugated antibodies procured from commercial sources, or investigator-derived monoclonal (mAb) or polyclonal (pAb) antibodies detected by indirect labeling.
• Assessment of early and intermediate CD4 or CD8 T cell activation in vitro by the expression of CD69 and CD25.
• Enumeration of lymphocytes expressing intracellular cytokines after stimulation in vitro with mitogens or antigens in the presence of brefeldin A or monesin, permeabilization, and labeling with fluorescent anti-cytokine antibodies.
• Discrimination of Th1/Th2 cells by the expression of CXCR3 vs CCR3, or CXCR3 vs CCR4.
• Enumeration of lymphocyte subsets in various stages of the cell cycle by surface phenotyping in conjunction with DNA dyes such as propidium iodide (PI) and SYTOX stains, or simultaneous detection of the Ki67 nuclear antigen.
• Quantitation  and simultaneous immunophenotyping of proliferating cells following in vitro stimulation by the detection of BrDU uptake or dilution of CFSE internsity.
• Quantitation of lymphocytes and other cells undergoing apoptosis by detection of Annexin V, enumeration  of hypodiploid cells after staining with propidium iodide, or detection of DNA fragmentation by the terminal deoxynucleotidyl transferase (TUNEL) technique.
• Detection and enumeration of foxp3 expression in Treg cells.
• MPA measurements of user-configured human or murine cytokine panels using  Luminex xMAP technology.
• Multiplex qRT-PCR on flow-sorted rare cells (in development).

Rate on FACSCalibur:	$75.00 per hour
Rate on FACSCalibur, hands-on:	$30.00 per hour
Rate on FACSAria II:	$100.00 per hour
Rate on FACSAria II:	$75.00 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.

Human Cytokine Analyses:


Cost per specimen (human analytes only) assayed in duplicate:	$20.00 for 2 plexes plus $5.00 for each additional plex.
Minimum fee:	$300.00 per assay.
Rate on Luminex, hands-on:	$100.00 per 96-well plate



Non-human Cytokine analyses:


Cost per specimen in duplicate:	$300.00 per plate for ≤ 40 samples
Cost per specimen run singly:	$300.00 per plate for ≤ 40 samples, plus $5.00 per sample above 40 samples

A 10% surcharge will apply to FACS Analyses and Cell Sorting, and Multiplex (MPA) Detection costs for Non-NYUSM Affiliated Users.

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.


Off-line computers running CellQuest, ModFit, Diva, and FlowJo are available on-site, at no charge, to users who process their own data. Those who wish to sign out one of many portable computers are charged $25 per night during the week or $25 over the weekend.