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Flow Cytometry and Cell Sorting Resource Laboratory (Penn)

Director: Moore, Jonni S., Ph.D.

Summary:

The Flow Cytometry and Cell Sorting Resource Laboratory is currently recognized as one of the largest and most comprehensive flow cytometry laboratories in the US. In 2010 it was designated a laboratory of exceptional merit by the National Cancer Institute. Using state-of-the-art technology, the resource provides a broad array of, instrumentation, support, education and consultation to the research community at the University of Pennsylvania. A wide variety of cell sorting applications are supported, from high-speed multicolor (up to 14 colors) cell sorting to low-speed, large nozzle, improved viability sorting. Additionally, a wide variety of cell analysis services (up to 20 parameters) are offered, from traditional analog, easier to use tabletop analyzers to many-laser, many-color, high-speed, fully-digital modern instrumentation. Currently the facility offers 6 cell sorters and 19 analytical instruments. A very active training and consultation program is in place to support these activities. The Scientific Director, Dr. Jonni Moore, and the Technical Director, each have over 25 years experience in the field of cytomics. Researchers at the University of Pennsylvania are increasingly engaged in research projects that require 8-plus-parameter cell sorting of infectious cells and primary human tissues. Investigators using the Flow Cytometry and Cell Sorting Shared Resource have access to virtually any type of cytometric services required for a vast array of applications.

Affiliations:

People:

Resources:

Instruments

  • Amnis ImageStream ( Flow cytometer )

    4 lasers: 405nm, 488nm, 642nm, 785nm
    Up to 10 fluorescent colors
    Has an extended depth of field
    Associated workstation to analyze images

  • BC Vi-CELL XR cell viability analyzer ( Automatic cytometer )

    The Vi-CELL XR utilizes the widely accepted tryphan blue dye exclusion method to determine cellular viability.

  • BD Accuri C6 analyzer ( Flow cytometer analyzer )

    Features:
    • Two lasers:
    —> blue (488nm)
    —> red (637nm)
    • Four fluorescent parameters

  • BD FACS Jazz in biosafety cabinet ( Flow cytometer sorter )

    Biohazardous cell sorter in biosafety cabinet
    jet-in-air
    3 lasers: Violet, Blue, Yellow
    Up to 6 color parameters

  • BD FACSAria II SORP sorter ( Flow cytometer sorter )

    • High or low speed sorter
    • Non-infectious cell sorting

    The core has two Aria sorters.

    «««««« Aria A »»»»»»
    • Three lasers:
    —> violet (407nm)
    —> blue (488nm)
    —> red (640nm)
    • 10 parameter detection

    «««««« Aria B »»»»»»
    • Four lasers:
    —> violet (407nm)
    —> blue (488nm)
    —> green (532nm)
    —> red (640nm)
    • 14 parameter detection

    Aria A & B features:
    • Four-way sorting
    • Single cell clone corting
    • Adjustable filters

  • BD FACSAria II SORP sorter in biosafety cabinet ( Flow cytometer sorter )

    • High or low speed sorter

    • Four lasers
    —> violet (407nm)
    —> blue (488nm)
    —> green (532nm)
    —> red (640nm)

    • 14 parameter detection

    • Features:
    —> can sort potentially infectious samples
    —> four-way sorting
    —> single-cell clone sorting
    —> adjustable filters

  • BD FACSCalibur analyzer ( Flow cytometer analyzer )

    The core has several FACSCalibur analyzers.

    Features:
    • Two lasers:
    —> blue (488nm)
    —> red (637nm)
    • 4 fluorescent parameters

  • BD FACSCanto analyzer ( Flow cytometer analyzer )

    The core has several Canto analyzers.

    Features:
    • High speed analyzer
    • Three lasers:
    —> violet (407nm)
    —> blue (488nm)
    —> red (633nm)
    • 11 color

  • BD FACSCanto II ( Flow cytometer analyzer )

    • High speed analyzer
    • Three lasers:
    —> violet (407nm)
    —> blue (488nm)
    —> red (633nm)
    • 8 color

  • BD FACSVantage SE sorter with FACSDiVa Option ( Flow cytometer sorter )

    • High or low speed sorter
    • Non-infectious cell sorting

    The flow core contains two DiVa Sorters.

    «««««« DiVa 206 »»»»»»
    • Three lasers:
    —> violet (407nm)
    —> blue (488nm)
    —> red (647nm)
    • 8 fluorescent parameters
    • Location: 206 John Morgan Building

    «««««« DiVa 297 »»»»»»
    • Three lasers:
    —> UV (355nm)
    —> blue (488nm)
    —> red (647nm)
    • 8 fluorescent parameters
    • Location: 297 John Morgan Building

    DiVa 206 & 297 features:
    • TurboSort option
    • Four-way sorting
    • Single cell clone sorting
    • Capability to change laser wavelengths to allow assays that are not optimized on table-top analyzers
    • Adjustable filters

  • BD Influx in biosafety cabinet ( Flow cytometer sorter )

    Biohazardous cell sorter in biosafety cabinet
    jet-in-air
    4 lasers: Violet, Blue, Green, Red
    Up to 13 color parameters

  • BD LSR II ( Flow cytometer analyzer )

    multi-laser
    multi-color
    fully-digital analyzer

  • BD LSRFortessa ( Flow cytometer analyzer )

    multi-laser
    multi-color
    fully-digital analyzer

  • Flow cytometry data analysis workstation ( Data analysis workstation )

    For analysis of flow cytometry data.

    Location: 220 John Morgan Building

Services

  • BSL2 Cell Sorting ( Access service )

    "We offer many instrumentation options for sorting biohazardous cells:

    BD FACS Aria II SORP - 2 instruments located in BRB II/III
    The Aria sorters are configured for 14-color detection with 4 solid-state lasers. They are housed within a custom-designed BioProtect IV hood which will ensure protection from aerosols that may contain potentially infectious materials such as HIV, SIV, and LCMV.

    BD Influx - 1 instrument located in Smilow Center for Translational Research
    The Influx combines traditional jet-in-air fluidics with analog log amps, but using higher bit rate ADCs and new Sortware software. Equipped with 4 lasers and 13 detectors, it can also sort six populations simultaneously.

    BD FACS Jazz - 1 instrument located in John Morgan Building 297/298
    The Jazz is a sorter conveniently located in our main lab and equipped to handle human samples. It is the newest cell sorter, equipped with 3 lasers and able to sort up to 6 colors."

  • BSL2 with Enhanced Precautions Special Training ( Training service )

    Training on how to use the BSL2 hood.

  • Cell sorting and analysis service ( Material processing service )

    "The staff performs special cell sorting applications on the BD FACS Aria II, BD FACSVantage SE, BD Influx and BD FACS Jazz."

  • FACSAria training ( Training service )

    Investigator must demonstrate proficiency using DiVa software.

  • Flow cytometer analyzer access ( Access service )

    "The facility offers training for investigator performed analysis on all bench-top analyzers."

  • Flow cytometry consultation service ( Support service )

    "Besides instrument training, the Directors and staff are always available for consultation on experimental design and to provide support for data acquisition for applications such as: immunophenotyping, rare event studies, functional probes, DNA cell cycle and apoptosis."

  • Flow cytometry data analysis ( Access service )

    Flow cytometry data are analyzed on the analysis workstation.

  • Flow cytometry data analysis services ( Data analysis service )

    "The Facility can advise and/or collaborate on implementing advanced data analysis methods for large-scale experiments in conjunction with Dr. Wade Rogers. Investigators may also access commercial data analysis software either by licensing through the Facility (for a quarterly fee) or by using a workstation maintained by the Facility."

  • ImageStream Analysis on ImageStream Work Station ( Data analysis service )

    Investigator performed or staff performed

  • ImageStream data acquisition ( Material analysis service )

    Investigator performed or staff performed

  • ImageStream Training ( Training service )

    Acquisition & Analysis Package

  • Non-infectious cell sorting access ( Access service )

    After training, the investigator can carry out cell sorting using the BDFACSAria II SORP.

  • Training: Lectures and Analyzer Instrument Classes ( Training service )

    "The lecture series is free of charge to any interested in attending. Introduction to Flow Cytometry lecture is required for everyone planning to do investigator-performed services."

Software

  • BD FACSDiva ( Software )

    "BD FACSDiva™ software is a collection of rich tools for flow cytometer and application setup, data acquisition, and analysis that help streamline flow cytometry workflow for today's busy laboratory. BD FACSDiva software offers built-in features for consistent and maintainable daily cytometer setup that supports reproducible data acquisition and reliable results."

  • CellQuest Pro ( Software )

    "BD CellQuest Pro software allows you to acquire and analyze data from your flow cytometer on a Macintosh® computer. Working in the BD CellQuest Pro Experiment document window, you can create several types of plots, including multicolor contour plots and overlaid histograms, and you can generate statistics for dot plots, histograms, density plots, 3D plots, and contour plots. The versatile graphics tool palette helps you produce presentation-quality documents. You can also save everything in an Experiment document, including plots, regions, gates, markers, statistics, calculated expressions, Browser contents, annotations, and color preferences, and then restore it all later."

  • FCS Express ( Software )

    "FCS Express can rapidly analyze flow cytometry data, with industry leading flow cytometry tools, plots, gates, and direct export to powerpoint (ppt), pdf, excel, word, and microsoft office."

  • FlowJo ( Software )

    "FlowJo should be the first step in the process of analyzing flow cytometry data. FlowJo can analyze data generated by any flow cytometer from any manufacturer. FlowJo has a number of different analysis platforms that let you not only perform standard analyses such as gating and statistics, but also specialized analyses such as DNA/Cell Cycle, Kinetics (Calcium flux), Proliferation, Calibration, and Statistical Comparison. You may find that FlowJo's sophisticated tools for generating output (graphical or tabular) are sufficient for you to generate publication-quality material.... but if not, FlowJo makes it easy for you to copy any graphs, statistics, or other information into other programs for further analysis and presentation."

  • Kaluza ( Software )

    "Kaluza is a revolutionary flow cytometry analysis software solution designed for high content data. Kaluza provides visualization tools, speed and an innovative simplicity to the flow community. With features designed to make your life easier in the lab, Kaluza aims to provide a greater understanding of the data generated by modern cytometers. Kaluza employs technology that allows for real time analysis of high content files allowing you to spend your valuable time on discovery."

  • ModFit LT ( Software )

    "ModFit LT is the only software that provides open-ended modeling of flow cytometry histograms. Models for use with cell-tracking dye studies and synchronized cell lines are built right into the software. It is, simply, the industry standard."

  • WinList ( Software )

    "Load FCS files from today's most popular instruments and do your FACS data analysis with a full set of region tools and gates. Easily design professional reports that update automatically. Create ratio parameters and user-defined equations, and apply fluorescence compensation to extend your analysis capabilities. Immunophenotyping, histogram subtractions, leukemia-lymphoma analysis, rare-event analysis, color gating - it's all possible with WinList."


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Last updated: 2015-01-30T15:19:10.702-05:00

Copyright © 2016 by the President and Fellows of Harvard College
The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016