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Flow Cytometry Core Laboratory (CHOP)

Director: Tuluc, Florin, M.D., Ph.D.

Summary:

The Flow Cytometry Core Laboratory provides access to state of the art instrumentation and professional flow cytometry services to members of the research community of The Children's Hospital of Philadelphia and University of Pennsylvania; investigators from outside the campus are welcome to our facility. The lab has space on the 12th floor of the Leonard and Madlyn Abramson Pediatric Research Center and on the fourth floor in Colket Translational Research Center. The staff has the required expertise for performing a variety of flow cytometry applications, including but not limited to sample processing for surface and intracellular staining, functional assays, complex multi-color flow cytometry analyses and cell sorting.

Affiliations:

People:

Resources:

Instruments

  • Analysis workstation with flow cytometry software ( Computer workstation )

    FlowJo, FACSDiva 8, CFlow Plus (Accuri)

  • BC MoFlo Astrios encased in a Class 2 Biosafety cabinet ( Flow cytometer sorter )

    This sorter has four lasers, with the following filters:

    • Laser: Violet (405 nm) •
    Filter: 448/59, 525/50, 670/30, 710/45

    • Laser: Blue (488 nm) •
    Filter: 513/26, 695/40

    • Laser: Yellow-Green (561 nm) •
    Filter: 579/16, 620/29, 644/22, 795/70

    • Laser: Red (640 nm) •
    Filter: 671/30, 722/44, 795/70

  • BD Accuri C6 analyzer ( Flow cytometer analyzer )

    This analyzer has two lasers and four fluorescent channel detection.

    • Laser: Blue (488 nm) •
    Filter: 530/30, 585/420, 670LP

    • Laser: Red (640 nm) •
    Filter: 675/25

  • BD FACSCalibur analyzer ( Flow cytometer analyzer )

    This analyzer has two lasers and four fluorescent channel detection.

    • Laser: Blue (488 nm) •
    Filter: 530/30, 585/42, 670LP

    • Laser: Red (635 nm) •
    Filter: 661/16

  • BD FACSJazz encased in a Class 2 Biosafety cabinet ( Flow cytometer sorter )

    This sorter has two lasers, with the following filters:

    • Laser: Blue (488 nm) •
    Filter: 530/40, 585/29, 692/42, 750LP

    • Laser: Red (635 nm) •
    Filter: 660/20, 750LP

  • BD FACSLSRII analyzer ( Flow cytometer analyzer )

    This analyzer has three lasers, with the following filters:

    • Laser: UV (355 nm) •
    Filter: 450/50, 530/30

    • Laser: Blue (488 nm) •
    Filter: 530/30, 585/42, 695/40, 780/60

    • Laser: Red (640 nm) •
    Filter: 670/14, 780/60

  • BD LSRFortessa analyzer ( Flow cytometer analyzer )

    This analyzer has five lasers, with the following filters:

    • Laser: UV (355 nm) •
    Filter: 379/28, 450/50

    • Laser: Violet (405 nm) •
    Filter: 450/50, 525/50, 610/20, 655/8, 710/50, 780/60

    • Laser: Blue (488 nm) •
    Filter: 530/30, 695/40

    • Laser: Yellow-Green (561 nm) •
    Filter: 582/15, 610/20, 660/20, 710/50, 780/60

    • Laser: Red (640 nm) •
    Filter: 670/14, 710/50, 780/60

  • Countess II FL ( Automatic cytometer )

    "• Automatic hemocytometer based on bright field and fluorescence imaging.
    • Count live / dead cells and measure average cell size in as little as 10 seconds.
    • Reusable slide available in the flow lab.
    • Compatible single-use slides: Invitrogen cat.# C10228; BioRad cat.# 1450011"

  • Evos FL Auto ( Microscope )

    "Evos is a fully-automated, inverted, multi-channel fluorescence / transmitted light imaging system.
    • Long working distance objectives: 1.25x, 4x, 10x, 20x, 40x (objectives are parfocal and parcentric).
    • Color camera for bright field and phase imaging.
    • Monochrome camera for fluorescence detecFon.
    • Motorized 3D stage.
    • Built-in soGware for counFng cells, scanning slides or plates, Fme-lapse imaging, Z stacks.
    • Light cubes may be swapped between Evos and the Countess."

  • Union Biometrica (COPAS) BioSorter - large particle sorter (up to 1000um) ( Flow cytometer sorter )

    This sorter has two lasers, with the following filters:

    • Laser: Blue (488 nm) •
    Filters: 510/23, 543/22

    • Laser: Yellow-Green (561 nm) •
    Filter: 615/24

Services

  • BSL2 biohazardous cell sorting ( Material processing service )

    Core staff carry out cell sorting of potentially infectious samples in a biosafety level 2 containment area.

    Samples requiring containment above BSL2 will not be sorted.

  • Flow cytometer access ( Access service )

    Access to instruments can be assisted or unassisted.

  • Flow cytometer training ( Training service )

    Training on operating the instruments existing in the core is provided upon request.

  • Flow cytometry consultation service ( Support service )

    Consult the core for:

    • requesting flow cytometry assays for basic and clinical research studies
    • assistance with developing functional flow cytometry assays
    • flow cytometry data analysis assistance
    • requesting training

    For additional details and to receive quotations for specific services contact Dr. Florin Tuluc at tuluc@email.chop.edu.

  • Flow cytometry data analysis service ( Data analysis service )

    Assistance with analysis of existing flow cytometry data is provided.

  • Flow cytometry online scheduling service ( Support service )

    Online scheduling for all instruments

  • FlowJo workstation access ( Access service )

    Users have access to two workstations (PC and Mac) running FlowJo software for data analysis.

  • Functional flow cytometry and bead-based assay service ( Material analysis service )

    Functional assays are offered on a fee-for-service basis for studies on animal and human samples.

    Examples of functional assays:
    • Apoptosis
    • Cell proliferation and viability
    • Cell cycle
    • Mitochondria membrane potential and generation of reactive oxygen species; mitochondrial mass
    • Intracellular calcium assays by flow cytometry
    • Phagocytosis
    • Detection of intracellular proteins (including cytokines and phosphorylated proteins using phospho-specific antibodies)
    • Cytometric Bead Arrays (CBA) for standard cytokines (Th1, Th2, Th17) or Flex CBA for human or mouse cytokines
    • Natural killer cells cytotoxicity assay

    Assistance with developing functional flow cytometry assays is also provided.

  • Non-infectious cell sorting ( Material processing service )

    Core staff carry out cell sorting for pathogen-free samples in a biosafety level 1 containment area.

  • Sample processing and flow cytometry analysis service ( Material analysis service )

    Sample processing (surface and intracellular staining) and flow cytometry analyses are offered on a fee-for-service basis.

    Most common analyses include immunophenotyping of human blood samples from clinical studies:
    • blood cell subsets (T, B, NK cells, monocytes)
    • T cells (naïve, memory)
    • immunoregulatory T cells (Tregs), follicular T-helper cells (Tfh)
    • expression of inhibitory/activating NK receptors
    • circulating progenitor cells
    • other phenotypes as requested by investigator.

    Other flow cytometry assays are available upon request.

Software

  • CFlow Plus ( Software )

    Accuri C6 software

  • FACSDiva ( Software )

    FACSDiva is the software used for data acquisition and analysis on LSR-type instruments (LSRFortessa, LSRII). The software is streamlined for generating reports directly from the platform used to acquire the flow cytometry data.

  • FlowJo ( Software )

    FlowJo can be used to analyze data generated on most flow cytometers and it has different platforms for standard, basic analyses such as gating and statistics, but also for specialized analyses, such as DNA/Cell Cycle, Kinetics (calcium flux), Proliferation, Calibration, and Statistical Comparison. FlowJo's tools are designed for generating graphic and tabular output sufficient for preparing publication-quality materials. Graphs, statistics, or other information can be easily copied and pasted from FlowJo into other programs for further analysis and presentation.


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Last updated: 2015-07-02T14:30:50.278-04:00

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