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Forsyth Multiplex Core

Director: Stephens, Danielle, M.S.

Location: Forsyth Institute,, 245 First St., Cambridge, MA 02142

Summary:

Bead-based multiplex arrays are commonly used for the detection of various biomarkers and proteins. This technique uses microsphere beads, coated with monoclonal antibodies against specific proteins, to simultaneously measure multiple analyte concentrations within a single small volume sample in various sample types.


  • body fluids (serum, saliva, plasma, gingival crevicular fluid, wound fluid, cerebral spinal fluid, joint fluid, etc)


  • cell and tissue extracts


  • cell culture supernatants


  • target species including human, mouse, rat, non-human primate, canine, porcine, etc.


Luminex® and their partners have developed many applications using the XMAP® Technology platform, including, but not limited to, the following fields of life science research:


































Cancer Markers Cardiovascular
Cell Signaling Cellular Metabolism
Immunology Inflammation
Gene Expression Profiling Genotyping
Endocrinology Isotyping
Matrix Metalloproteinases Metabolic Endocrinology
Neurobiology Transcription Factors
Toxicity


The Forsyth Multiplex Core, established in 2013, offers three main platforms:

  • Luminex® 200™

  • FLEXMAP 3D™

  • AYOXXA Lunaris™ Technology


All platforms operate 96-well plate based assays. FLEXMAP 3D™ and AYOXXA also offer 384-well plate capacity.
Plate capacity: up to 80 samples in singlet with up to 16 wells designated as standards for antigen-antibody based assays (for 96-well format).

Capabilities

  • Multiplex protein detection using antibody-based assays of up to 100 analytes or 500 analytes in a single microplate well using the Luminex® 200™ or FLEXMAP 3D™, respectively.


    • Using 5-50 µl of sample volume, pg/mL levels of hundreds of protein biomarkers are detectable.


  • Ability to custom coat beads with various mammalian and non-mammalian molecules such as nucleic acids, receptor-ligands, and siRNA. These require custom-plexing, assay development/optimization, and additional pricing.

  • Both Luminex systems use BioPlex Manager, from Bio-Rad, for acquisition and computation of data.


    • Allows easy transfer of the data to excel.

    • In addition to the raw and computed data on fluorescence intensity and concentrations, standard curves, %CV, internal control samples, plate-to-plate variation, and standard deviation are only a few of the data output measures.

    People:

    Resources:

    Instruments

    • Automated pipette station (Integra) ( Multichannel pipette )

    • AYOXXA Lunaris™ Reader 96/384 ( Microplate reader )

    • Barcode Scanning Capabilities ( Barcode reader )

    • Bio-Rad Bio-Plex Pro™ Wash Station ( Microplate washer )

      The Bio-Plex Pro Wash Station eliminates manual wash steps from Bio-Plex Assays, making them as easy as ELISAs. The wash station is specifically designed to perform Bio-Plex Assay wash steps, but it is also compatible with any standard xMAP assay.

      Features and Benefits of the Bio-Plex Pro Wash Station

      Improved lab productivity
      Reliable and reproducible results
      Optimized on-board protocols
      The Bio-Plex Pro Wash Station incorporates a magnetic plate carrier for reliable, hands-free wash steps on magnetic bead-based assays (Bio-Plex Pro and MagPlex Assays).

      Preset wash programs have been optimized for Bio-Plex Assays. Using the Bio-Plex Pro Wash Station in an assay workflow reduces manual intervention and decreases variability between independent experiments.

    • FLEXMAP 3D™ ( Microarray platform )

      The next generation multiplexing platform based on xMAP technology. Expanded multiplexing capability, faster time to results, and automation capability make it the platform of choice for high-throughput testing for nucleic acid and protein applications.
      Measure up to 500 unique analytes in a single sample
      96- and 384-well plate capability
      Plate read times twice as fast as the Bio-Plex/Luminex 200 system
      LIS-compatible software
      Compatible with the magnetic and nonmagnetic assays
      Bio-Plex Manager™ software version 6.0 for data analysis"

    • Luminex LS 200 Bead Array Instrument ( Bead array reader )

    • Temperature-sensitive storage conditions for +4°C, -20°C, and -80°C (short term storage) ( Cold room )

    • ThermoFisher KingFisher Flex ( Nucleic acid extraction/purification instrument )

      Used for sequential multiplex analysis.
      Features:
      - High-throughput purification of 24-96 samples per run
      - Easy to install and run (ready to run in 10 minutes or less)
      - Choose from two plate formats for wide volume range (20–5,000 µL)
      - Download existing protocols from library, or easily customize protocols using Thermo Scientific BindIt Software
      - Simple workflow

    Services

    • Full service analysis ( Material analysis service )

      Full-service analysis: researchers provide a list of desired biomarkers to study. The Core will provide the best configuration of panels commercially available to analyze the maximum number of desired markers. Core will provide a quote for services. Researcher sends samples to the Forsyth Institute for temperature-controlled storage until time of analysis. The Core orders and receives all kits and materials necessary for analysis. Analysis is performed entirely by Core personnel and data is exported and provided as an excel spreadsheet with desired data sets. No statistical data analysis is performed. Minor data organization can be performed by the Core upon request.

    • Read-only analysis ( Material analysis service )

      Researcher purchases panels needed for analysis, performs sample and kit preparations, simply brings the finished plate to the Forsyth Institute to be read on our equipment by Core personnel. Consultation for panel configuration can be performed by Core personnel upon request. Data is exported and provided as an excel spreadsheet with desired data sets. No statistical data analysis is performed. Minor data organization can be performed by the Core upon request.


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    Last updated: 2020-01-08T15:47:37.217-06:00

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