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Immune Monitoring and Flow Cytometry Resource (IMFC)

Director: Channon Smith, Jacqueline Y.

Summary:

DartLab is a Shared Resource that is used both by basic scientists and by clinicians carrying out mechanistic studies for clinical trials. Services are also available to non-Dartmouth clients. The consolidated Shared Resource moves beyond the simple provisioning of technical services to making complex experiments possible by providing scientific and technological support for implementing advanced, high-complexity assays in a cost-effective way, and by providing a mechanism for the acquisition of new methodologies.

We carry out a spectrum of immunoassays and can develop customized immunoassays to suit your studies. We provide dedicated instruments for your use including cell sorters, flow cytometers, an ELISPOT reader, a Bio-Plex array reader and a Sector Imager, an autoMACS and a Robosep.
Whole Blood Services

* Flow Cytometry
* Whole blood processing
* Cell Subset Enrichment
* Education
* Clinical Trial Studies

Affiliations:

People:

Resources:

Instruments

  • autoMACS cell sorter ( Cell separator )

    "The autoMACS Separator is a benchtop instrument for high-speed automated cell enrichment. Employing MACS Technology, the autoMACS Separator is designed for positive selection as well as depletion of magnetically labeled cells."

  • BD 4-Color FACSCalibur cytometer ( Flow cytometer analyzer )

  • BD 6-Color FACSCanto cytometer ( Flow cytometer analyzer )

  • BD FACSAria I cell sorter ( Flow cytometer sorter )

  • Beckman Coulter 10-color Gallios flow cytometer ( Flow cytometer )

    "The Gallios Flow Cytometer is a research system that delivers analytical excellence by coupling extraordinary sensitivity, resolution and dynamic range with high-speed data collection.

    Patented electronics provide accurate and efficient digital signal processing at high event rates over a wide dynamic range of fluorescence intensities. These benefits are packaged in a compact analyzer that delivers stable performance over long periods of time and across a wide range of operating temperatures.

    The Gallios houses up to three highly reliable, solid-state lasers in standard red and blue, with violet available as an option. Easily interchanged optical filters facilitate detection of a variety of dyes and wavelengths.

    An innovative forward-scatter detector provides up to three measurements of cell size and visualization of particles down to 0.404 µm in diameter. A side-scatter detector incorporates an independently focused, high-performance photodiode with electronic attenuation. Six fluorescence detectors provide simultaneous acquisition of up to six fluorescence signals. Up to four fluorescence detectors can be added to the system, enabling concurrent reading of up to 10 colors. Simultaneous measurements of integral, peak and width are available for all parameters, including scatter and fluorescence signals. A selection of up to 62 parameters can be processed per analysis, at acquisition rates of 25,000 events-per-second, with high yield. Three scalable configurations allow for the future addition of more lasers and fluorescence detectors as the applications for complex multi-color flow cytometry expand."

  • Bio-Plex Array Reader ( Arrayer )

    "Bio-Plex MCV plate IV — maintenance, calibration, and validation plate customized for hands-free startup, shutdown, and performance testing.

    Permits analysis of up to 100 biomolecules in a single sample.

    Calibration and validation kits — system tools for daily calibration of signal output and for monthly validation of primary performance parameters of the reader.

    High-throughput fluidics — delivers up to 20 L of sheath fluid (40 plates) without user intervention.

    Combines 2 lasers, fluidics, and real-time digital signal processing to distinguish up to 100 different color-coded bead sets, each representing a different analyte.

    Bio-Plex Manager software — controls the instrument, data acquisition and analysis for superior curve fitting, statistics reporting, and charting features, with no requirements to export or import files.

    Bio-Plex SNP Manager data analysis software — simple-to-use data analysis tool for nucleic acid applications."

  • KS ELISPOT reader ( ELISPOT microplate reader )

    "Comprises a stereomicroscope with zoom optics. If you want to analyze your ELISPOT samples quickly, acquire them using one image per well. In this case you can accurately detect spots with diameters down to 30 µm."

  • Miltenyi Biotec 7-Color MACSQuant analyzer ( Flow cytometer analyzer )

  • Miltenyi Biotec 8-color MACSQuant VYB flow cytometer ( Flow cytometer sorter )

    8-color MACSQuant VYB flow cytometer

    Equipped with three lasers, two scatter (FSC, SSC) and up to eight fluorescent channels, the MACSQuant Analyzers are the ideal instruments for rapid, fully automated multicolor flow cytometry.
    Absolute cell counts are determined volumetrically, calculated automatically, and read out as events/μL for every sample and every population. Cell counts are available within less than a minute. There is no need for expensive counting beads or particles.

  • Robosep cell separator ( Cell separator )

    "RoboSep offers walk-away automation of immunomagnetic cell isolation by performing all EasySep cell labeling and magnetic separation steps."

  • SECTOR Imager 2400 reader ( Microplate reader )

    "The SECTOR Imager 2400 reader is the newest member of the award-winning SECTOR Imager product line. The SECTOR Imager 2400 reader uses ultra-low noise charge-coupled device (CCD) camera with custom-designed telecentric lenses for rapid detection in all MULTI-ARRAY® or MULTI-SPOT® plates. It is ideal for applications requiring high information content and high throughput. Whether you are multiplexing biomarkers and cytokines or screening kinases, ubiquitin ligases, integrins or GPCRs, you will appreciate the simple no-wash formats and minimal consumption of reagents.

    The SECTOR Imager 2400 reader offers all the capabilities of the SECTOR Imager 6000 reader: high sensitivity for single or multiple analyte assays and other complex biological targets. The SECTOR instrument is a medium throughput imaging detection system, capable of multiplexing in all spot formats and reads 96- and 384-well plates."

Protocols

  • BioPlex protocol ( Protocol )

  • Cell surface staining ( Protocol )

    Cell Surface Staining

    STAINING OF CELLS WITH MONOCLONAL ANTIBODIES:
    ANALYSIS OF EXTRA-CELLULAR MEMBRANE PROTEINS BY FLOW CYTOMETRY

    From Givan, A.L. (2000), chapter in In Living Color: Protocols in Flow Cytometry and Cell Sorting (R. Diamond and S. DeMaggio, eds). Springer, Berlin, pp 142-164.

    Before staining cells, you should know about the CRITICAL ASPECTS of staining that are important for success and about the SOLUTIONS you will need.

  • MilliPlex protocol ( Protocol )

Services

  • Cell separation service ( Material production service )

  • Cell subset enrichment service ( Material production service )

  • Cell surface staining ( Material processing service )

    "Technicians available to stain with monoclonal antibodies and analyze cells by flow cytometry. In addition to the instrument time there is a fee for the technician time and reagent expense."

  • CFSE lymphoproliferation analysis service ( Material analysis service )

    "CFSE is used to intracellularly label proliferating T cells. The fluorescent intensity can be detected by flow cytometry and is used to measure proliferation and mitotic activity."

  • ELISPOT service ( Material analysis service )

  • Flow cytometry assay service ( Material analysis service )

  • Flow Cytometry Clinics ( Training service )

    Drop in Flow Cytometry Clinics are offered at the beginning of the Fall and Spring

    The Flow Core Director will be available in the Flow Core computer room from 2-5pm to assist you with planning multi-color experiments, using flow cytometry software, using instruments, or analysing your data."

  • Flow cytometry service ( Material analysis service )

  • Multi-plex cytokine assay service ( Material analysis service )

    "The multiplex cytokine assay format differs from conventional ELISA in one significant way—the multiplex capture antibody is attached to a bead whereas the ELISA capture antibody is attached to the microplate well. The technology uses 5.6 micron polystyrene or magnetic microspheres, which are internally dyed with red and infrared fluorophores of differing intensities. Each bead is given a unique number, or bead region, allowing differentiation of one bead from another. Beads covalently bound to different antibodies can be mixed in the same assay, utilizing a 96-well microplate format."

  • Regulatory T cell conversion service ( Material modification service )

  • Whole blood processing service ( Material processing service )

Software

  • FlowJo ( Software )

    Flow cytometry data and statistical analysis software.

  • FlowLogic ( Software )

    It combines flow cytometry data analysis, graphing, statistics and reporting. Flowlogic also offers a data array analysis function visualized as a plate. This is useful for fast analysis of large data sets. It also provides unique display options for all types of analyses and the easy generation of heat maps.

  • FreezerPro ( Software )

    "FreezerPro 2011 is a frozen sample management solution. It is a web-based application that enables users to know precisely where a frozen sample is located. Features include automatic alerts of low number of sample aliquots, sample expiration date, sample volume or freeze-thaw count along with powerful reporting."

  • GemStone ( Software )

    "Flow cytometry software for analysis of high-dimensional, flow cytometry data. Based on patented Probability State Modeling technology."

  • Kaluza­ Flow Analysis Software ( 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." – Kaluza Product Page Kaluza is a flow cytometry analysis software suite produced by Beckman Coulter. The software is designed to work seamlessly with the LMD files created by their Gallios cytometer—which, due to their 32-bit format, tend to have scaling issues when analyzed in FlowJo—but is also capable of analyzing FCS files of version 3.0 or later.

  • ModFit LT ( Software )

    "Versatile DNA analysis software featuring AutoDebris, AutoAggregates, AutoLinearity, and exceptional batch processing. ModFit LT 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."

  • MSD DISCOVERY WORKBENCH Software ( Software )

    "MSD's DISCOVERY WORKBENCH® software provides real-time data visualization for multiplex assays and allow users to create data files for easy export."

  • SECTOR Imager 2400 software ( Software )

    - On-board data storage in secure database
    - Real-time data visualization and processing tools
    - Custom data export for integration with external databases
    - Optional robotic integration tools

  • Winlist ( Software )

    "Flow cytometry analysis software, Winlist allows one to analyze your data 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."


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Last updated: 2014-10-02T15:57:29.514-04:00

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