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Cytomics platform

The cytometry platform incorporates the latest technologies in the field of flow cytometry, applied to both the analysis and separation of cells. Cytometric techniques make it possible to quickly and easily analyze and sort a significant number of cells with high sensitivity and objectivity. It is also possible to obtain qualitative and quantitative measurements and to define complex cell subpopulations from the study of a set of parameters defined for each specific cell.

During the last few years, "full-spectrum" cytometry has been developed, which allows more individual fluorochromes to be resolved by collecting and processing the data as complete spectrums. Thus, it is possible to design immunophenotyping panels with more than 40 fluorescent parameters, as well as having other important advantages over conventional cytometry. Recently, separator equipment incorporating this technology has been developed, such as the Aurora CS which is available at UAT.

Services:

  • Cell analysis by flow cytometry.
  • Fluorescence-activated cell segregation (FACS).
  • Analysis of multiplexed genomic and proteomic biomarkers.

We also offer training in the self-use of the equipment, advice on cytometry techniques, protocol development, panel design, support in the interpretation of results and data analysis.

You can find the "welcome handbook" to the cytometry platform here.

Applications:

  • Flow cytometry:
    • Immunophenotyping from the surface and intracellular antigen labeling.
    • Nucleic acid analysis (ploidy, cell cycle).
    • Functional analysis (e.g. enzyme activity, cell integrity, membrane permeability, calcium fluxes).
    • Viability, proliferation, apoptosis assays.
    • Assessment of transfection efficiency (e.g., GFP, mCherry, etc.).
    • Tiny particle analysis.
  • Cell separation:
    • Isolation of well-defined cell populations from heterogeneous samples (up to 6 populations at a time
    • Isolation of rare populations (e.g., CTCs, CSCs).
    • Isolation of fluorescently labeled cells (e.g., transfected cells).
    • Single-cell isolation in 96-well plates.
  • Multiplexed genomic and proteomic biomarker analysis. Multiplex tests with Luminex® technology are designed for the simultaneous detection and quantification of several secreted proteins such as cytokines, chemokines or growth factors, or expressed genes. This high-throughput technology generates results comparable to those of conventional methods such as ELISA, with the use of a reduced sample volume (25 µl of plasma or 50 µl of cell culture supernatant) in a 96-well plate format.

If you have any questions, please contact us at citometria@vhir.org.

Equipment:

  • Flow cytometers:
    • FacsCalibur cell analyzer (Becton Dickinson): currently has only one blue laser, which allows detection of two fluorochromes in this spectral range (plus FSC and SSC). 
    • LSRFortessa cell analyzer (BD): 4 lasers (blue, red, violet and yellow-green) for parallel detection of up to 16 fluorochromes (plus FSC and SSC). Acquisition in tube and HTS format (96- and 384-well plates).
  • Cell separators:
    • FACSAria high-speed cell separator (BD): 4 lasers (blue, red, violet and yellow-green) for detection of up to 13 fluorochromes (plus FSC and SSC). Separation in 4 lanes (0.5 to 15ml collection tubes) and in 6- to 96-well plates, including single cell separation in 96-well plate. Cytometer configurations can be found here.
    • Cell separator with Aurora CS spectral technology (Cytek): this is a "full-spectrum" technology equipment, which detects and processes the spectral fingerprint of each fluorochrome, providing multiple advantages over conventional cytrometry: increase in the number of fluorochromes that can be detected in the same panel, extraction of auto-fluorescence treating it as an independent parameter and distinction between fluorochromes with similar emission maximums.
      • This separator is equipped with five lasers (near-UV, violet, blue, yellow-green and red) plus an array of fluorescence detectors for each laser (capturing from 365 to 829 nm), plus FSC and two SSCs (one of them optimized for small particles), with a total of 67 detection channels. The instrument allows separation in up to 6 lanes or in 96-well plates, with the "index sorting" option. The instrument can also be used as an analyzer.
      • The equipment is incorporated into a class II, type 2A biosafety cabinet, which allows working with samples infected with pathogenic organisms within the previously mentioned biosafety level.
  • Multiplexed genomic and proteomic biomarker analysis
    • Luminex Magpix: this multiplex analysis kit performs up to 50 different tests in a single reaction volume, and allows a 96-well plate to be read in ≤ 60 minutes. Compatible with ProcartaPlex (immunoassays for secreted proteins), QuantiGene Plex (RNA transcripts) and other bead-based kits. Results can be exported and analyzed with the ProcartaPlex Analyst software.
  • Software packages for data analysis
    • We offer two off-line stations for data analysis with specific cytometry software:
      • FlowJo: 1 license installed locally on analysis station 1 (accessible on-site and remotely).
      • FCS-Express 7: 1 license, accessible in the cloud upon registration. Access must be requested from UAT staff.
      • SpectroFlo: software for initial acquisition and analysis of spectral cytometry data. 1 license installed locally on analysis station 4 (accessible on site and remotely).
      • OMIQ: software dedicated to the analysis of high dimensional data, especially spectral data. Accessible in the "cloud". It is necessary to request access to UAT staff.