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Flow Cytometry Core (JDC)

Directors: Serwold, Thomas F., Ph.D.; Wagers, Amy, Ph.D.

Location: Joslin Diabetes Center, One Joslin Place, Boston, MA 02115

Summary:

The Flow Cytometry Core Facility provides high speed cell sorting, multi-color fluorescence cell analysis and data interpretation to Joslin Investigators. The aim of this core facility is to provide a comprehensive service with high quality for scientific researchers. The staff at the facility works closely with investigators to develop new techniques to meet research needs. We welcome investigators interested in developing new modes of analysis.

The core is open to everyone. Different pricing for HSCI vs academic non-HSCI and non-academic institutions.

Affiliations:

People:

Resources:

Instruments

  • 3-Laser BL2 BD FACSAria II u ( Flow cytometer sorter )

  • 5-laser BD SORP ARIA ( Flow cytometer sorter )

    This instrument is capable of high speed and single cell sorting on up to 15 fluorescent parameters and is equipped with 5 lasers (355nm, 488nm, 405nm, 561nm, and 640nm). It is capable of four-way sorting into tubes or single cell sorting directly into plates or slides. This instrument is equipped with an Aerosol Containment System for the sorting of BSL-2 biohazardous samples.

  • BD FACSAria cell sorter ( Flow cytometer sorter )

    "The Aria is a high speed cell sorter (up to 22,000 cells/second) and analyzer (up to 70,000 cells/second). The Aria is a 3 laser system configured with a 488nm, 633nm and 407nm lasers. At the present time, we can detect 10 fluorescent parameters and 2 size parameters. The Aria will be upgraded to detect 13 fluorescent parameters in the future.

    2 and 4 way sorting is available. Sorting can be done into tubes of various sizes, plates and custom slides. Single cell sorting into plates is also available.

    Only the Aria is available for non-infectious human sorting or analysis with non-fixed cells."

  • BD LSR II analyzer ( Flow cytometer )

    "A 3-laser bench-top analyzer configured with 488nm, 633nm, 407nm lasers. The LSRII can detect 10 flourescent parameters and can be used independently by investigators (after training). For analysis of human cells, only fixed, non-infectious samples run on the LSRII."

  • BD LSRFortessa ( Flow cytometer analyzer )

    This instrument is equipped with five laser lines (445nm, 405nm, 488nm, 561nm, and 640nm) and 17 fluorescent detectors, and the HTS (High Throughput Sampler) system which will enable fully automated sample acquisition from 96- and 384-well microtiter plates. The current configuration allows for 2 colors off the 445nm laser, 3 colors off the blue laser including GFP and YFP detection, 5 colors off the yellow-green laser, 3 colors off the red, and 4 off of the violet.

  • Beckman/Coulter MoFlo cell sorter ( Flow cytometer sorter )

    "Another high speed cell sorter (up to 30,000 cells/second) and analyzer (up to 50,000 cells/second). The MoFlo is a 3 laser system configured with a 488nm, 633nm and UV lasers. The MoFlo can detect 6 fluorescent parameters and 2 size parameters. Because the MoFlo is equipped with a true UV laser, we are able to detect and sort SP (side populations) and monitor Calcium Flux in cells.

    2 and 4 way sorting is available. Sorting can be done into tubes of various sizes, plates and custom slides. Single cell sorting into plates is also available.

    Only fixed non-infectious human cell work can be performed on the MoFlo."

  • Cellometer Vision Trio Cellprofiler ( Instrument )

    The Cellometer Vision Trio Cellprofiler system combines bright field microscopy and fluorescence detection to determine total cell numbers, measure cell viability, and quantify GFP expression.

  • ELISPOT reader ( ELISPOT microplate reader )

    This analyzer is designed for scanning and evaluating a wide range of microtiter plate-based bioassays—in particular, ELISPOT assays. This instrument offers:
    • Uncompromised quality of analyzer components
    • Precise, fast, user-friendly scanning
    • Objective, reproducible counting results

  • Hemavet 950 FS ( Instrument )

    The Hemavet 950FS Multi Species Hematology System specializes in counting differential white blood cells (WBCs) with 24 parameters including total WBC, erythrocyte, lymphocyte, monocyte, neutrophil and other granulocyte subtypes, and platelet numbers.

  • Miltenyi autoMacs separator ( Cell separator )

    "A self-run bench-top instrument for high-speed automated cell sorting. It is designed for positive selection or negative depletion of magnetically labeled cells."

  • Miltenyi MACSQuant analyzer ( Flow cytometer analyzer )

  • Union Biometrica Large Particle BioSorter ( Flow cytometer sorter )

    "A newly developed large particle sorter that uses lower operating pressures (1-3 pse) and has a larger nozzle (250 microns) than conventional cell sorters. The BioSorter is suitable for analysis and sorting of large and/or fragile cells such as adipocytes, ES and EB cells. It is equipped with a 488 nm laser with 3 filter sets:

    * 527 nm to detect green dyes (FITC and Alexa 488)
    * 670 nm to detect red dyes such as PE
    * 700 nm or longer wavelengths to detect far-red dyes such as PerCp"

Services

  • Apoptosis analysis service ( Material analysis service )

    "Detection with Annexin V, PI."

  • Automatic cell counting ( Material analysis service )

    This service uses Cellometer and Hemavet for cell counting.

  • Calcium flux analysis service ( Material analysis service )

    "Calcium flux analysis is done using Indo-1."

  • Cell analysis ( Material analysis service )

    Uses the BD LSR-II, Miltenyi Biotec MACSQuant and BD LSRFortessa to perform fluorescent color analysis.

  • Cell cycle analysis service ( Material analysis service )

    "DNA content (PI, 7AAD, Hoechst33342), S-phase detection (BrdU), and CFSE for label dilution analysis of sequential cell divisions."

  • Cell sorting ( Material processing service )

    Service uses MoFlo Legacy, SORP BD FACSAria and BD FACSaria Hu for cell sorting.

  • Cell tracking service ( Material analysis service )

    "Cell tracking is done by using CFSE, SNARF, PKh26, PKH67 or PKH2."

  • Data analysis ( Data analysis service )

    For flow cytometry data analysis, the Core provides 3 FlowJo dongles that can be signed out from the core on a weekly basis.

  • EliSPot Reader ( Material analysis service )

    Scans and evaluates a wide range of microtiter plate-based bioassays, in particular, ELISPOT assays.

  • Fluorescent color analysis/sorting ( Material processing service )

    15 fluorescent color analysis/sorting on 5 laser BD SORP AriaII:
    lasers: 355, 407, 488, 561, and 640

    8 fluorescent color analysis/sorting on3 laser Beckman Coulter MoFlo:
    lasers: 355, 488 and 640

    12 fluorescent color analysis/sorting on 3 laser BD Aria IIu:
    lasers: 407, 488 and 640

    15 fluorescent color analysis on 4 laser BD LSRII
    laser: 407, 488, 561 and 640

    18 fluorescent color analysis on 5 laser BD LSRFortessa
    laser: 407,445, 488, 561 and 638

    7 fluorescent color analysis on 3 laser MACSQuant:
    lasers: 407, 488 and 647

  • Gene expression analysis service ( Material analysis service )

    "Gene expression analysis for reporters expressing GFP or GFP derivatives (YFP, CFP, etc.)"

  • Large object sorting ( Material processing service )

    Service uses the Complex Object Parametric Analyzer and Sorter (COPAS) BioSort system to sort objects ranging from 40-250 microns.

  • Magnetic cell enrichment ( Material processing service )

    This service uses AutoMACS to provide high yield enrichment of rare cell populations via magnetic selection, which helps to reduce per user sort times by enabling better pre-sort enrichment of target cells.

  • Single-cell sorting service ( Material production service )

    We provide single-cell sorting for cloning and DNA/RNA analysis.

Software

  • FlowJo ( Software )

    Flow cytometry data and statistical analysis software.

  • Summit ( Software )


Web Links:

Last updated: 2017-08-18T14:21:00.216-05:00

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The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016