1. SCIENCE
  2. CORE FACILITIES
  3. Flow Cytometry

Flow Cytometry

The Gladstone Flow Cytometry Core enables scientists to analyze or isolate cells based on their phenotypic or functional properties, and can be used in conjunction with other core technologies to form a robust and effective pipeline for cellular analysis.

Analysis of up to 64 and sorting of up to 18 fluorescent parameters is available from a wide variety of samples and cell types. Our staff is experienced in experimental and antibody panel design, practical operation of multiple flow cytometric platforms, and basic and advanced data analysis.

Services

Cell sorting for up to 18 fluorescent parameters with the following specifications:

  • 5 available lasers: UV (355nm), Violet (405nm), Blue (488nm), Yellow-Green (561nm), Red (640nm).
  • Functional marker sorting and analysis (e.g., phospho-flow, cell cycle, and mitochondrial assays).
  • Single-cell sorting into 96-well plates for cloning or further sequencing downstream analysis.

Sorting into the following receptacles:

  • 1.5ml Eppendorf tubes
  • 5ml Falcon tubes
  • 15ml Falcon tubes
  • Any well plate (12, 24, 48, 96 and 384)
  • Terasaki plate
  • Standard/frosted end slide

Aseptic sorting is available.

  • Analysis of up to 64 fluorescences in one tube.
  • Analysis of extra, intracellular and functional markers as well as fluorescent proteins.
  • High-throughput analysis from 96- and 384-well plates for all analyzers.
  • Imaging Cytometry for up to 6 fluorescences, or 4 fluorescences including brightfield and side scatter.
  • 20X, 40X and 60X magnification available.

Test whether the human induced pluripotent stem cells (hiPS cells) that you have received or grown express a panel of pluripotent markers and define their pluripotency status. Markers include Tra-1-60, Tra-1-81, SSEA-3, SSEA-4, CD30, CDH3, Nanog, Oct 3_4 and Sox-2, which you can choose from to run in your panel. Turnaround time is approximately 1 day and you’ll receive all raw data files at the conclusion of the experiment.

Contact the Flow Cytometry core for pricing.

Gladstone’s Flow Cytometry Core also offers sample processing, including staining and running of experimental samples. Data analysis services are also available.

Equipment

NanoFCM Analyzer

NanoFCM Analyzer

The NanoFCM Analyzer can be used for the multiparameter characterization, sizing and concentration of natural and synthetic nanoparticles (7-1000 nm) at the single-particle level, including detection of extracellular vesicles, mitochondria, bacteria, viruses, nanomedicine, nanomaterial and other nanoparticles.

Specifications:

  • Laser (blue, green, red)
  • One side-scatter channel
  • Two fluorescence channels
  • Up to 12,000 particles/min
  • Lower detection limit 7 nm (gold nanoparticles) to 40 nm (exosomes, viruses)
  • High detection limit (1 micron)

Practical and theoretical training is available for the Nano FCM, as well as experimental and data analysis support.

Fortessa X-20

Fortessa X-20

The Fortessa X-20 cell analyzer delivers high-performance, multicolor analysis and is configured with five lasers to detect up to 20 parameters (18 fluorescent and 2 scatter) simultaneously. An integrated High Throughput Sampler (HTS) provides rapid, fully automated sample acquisition from 96- and 384-well microtiter plates.

Practical and theoretical training is available for the Fortessa X-20, as well as experimental and data analysis support.

Cytek Aurora

Cytek Aurora

The Cytek Aurora is a spectral cytometry analyzer configured with four lasers (violet, blue, yellow/green, and red) to detect up to 48 parameters simultaneously. Other features include autofluorescence extraction, ability to determine fluorescences in one tube that are unable to be distinguished by regular flow cytometry, and intuitive software with transferable experimental workspaces.

Practical and theoretical training is available for the Aurora, as well as experimental and data analysis support.

Aurora configuration:

  • 16 violet parameters
  • 14 blue parameters
  • 10 yellow/green parameters
  • 8 red parameters
BD FACSAria Fusion Cell Sorter

BD FACSAria Fusion Cell Sorter

Two BD FACSAria Fusions cell sorters are configured to sort up to 20 parameters (18 fluorescence and 2 scatter). A choice of nozzles allows users sort a wide range of particle sizes. Nozzles are available in four sizes: 70, 85, 100, and 130 microns. Bulk, single cell and index sorting are available.

Practical and theoretical training is available for the Fusion, as well as experimental and data analysis support.

Attune NxT Flow Cytometer

Attune NxT Flow Cytometer

The Attune NxT Flow Cytometer is a compact, benchtop cell analyzer that has been configured with four lasers to run and analyze multicolor panels of up to 16 colors (14 fluorescence and 2 scatter). An integrated autosampler provides rapid, fully automated sample acquisition from 96- and 384-well microtiter plates.

Practical and theoretical training is available for the Attune, as well as experimental and data analysis support.

Amnis Imagestream X Mark II

Amnis Imagestream X Mark II

The Amnis Imagestream X Mark II is an imaging cytometer that combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy. The Imagestream comes equipped with 20, 40 and 60X magnification, as well as the ability to detect up to 6 fluorescent channels (or 4 plus brightfield and side scatter) using 4 lasers (405, 488, 561 and 635nm). Cells can be run at up to 5,000 events per second.

Applications for the Imagestream include:

  • Co-localization
  • Internalization
  • Cell cycle and mitosis
  • Cell signaling and interaction
  • Shape change and chemotaxis
  • Nuclear translocation

Practical and theoretical training is available, as well as experimental and data analysis support.

Training

We offer training on the equipment in the Flow Cytometry Core. Contact the core staff to start the practical training process.

To get started, you’ll need to:

You can also refer to additional resources to learn more about flow cytometry.

FAQs

You’ll first need to register for an iLab account. Once that’s complete, you should read, sign, and return the signature page of the general regulations document to the core by email.

Contact core staff if you have not used the core before.

The Fortessa X-20, Attune NxT, and Cytek Aurora, are capable of analyzing 96- and 384-well plates.

If you’d like to be trained on an analyzer or sorter, complete Gladstone’s Flow Core intro questionnaire to answer some questions about your experience, and email the core to schedule a date.

Any single-cell suspension where cells are between 500 nm and 20 μm can be run. Samples can be provided in 5-ml Falcon tubes, 15-ml Falcon tubes, Eppendorf tubes, multi-well plates, or slides.

Cells can be at a concentration of up to 5 million per mL in PBS or media, but serum should be limited to 2 percent serum, as more than this can affect the charge applied to the stream. You can also add HEPES (25 mM), EDTA (1 mM) or DNAse (10 U/ml) to avoid clumping. If you are concerned that your sample is too concentrated, bring in some media to dilute your sample. Filter your cells through a cell strainer or filter-top tube.

After being trained, you must demonstrate proficiency in cytometer operation, maintenance, and basic troubleshooting before being allowed to use the equipment independently or outside of operating hours.

The core saves a copy of the FCS files to an internal server.

Publications

Contact

Kevin Pastores
Assistant Core Director
Email

Members

Jane Srivastava
Scientific Resource Director
Email

Nicole Colligan
Research Technologist II
Email

Fees and Scheduling

For more information on our fees, contact the core.

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