Flow cytometry is a cell analysis technique that involves measuring the light scattering and fluorescence characteristics that cells possess as they are passed through a beam of light. Flow cytometry is used in the counting and classification of cells according to their morphological characteristics, presence of biomarkers, and in protein engineering. In flow cytometers, cells suspended in a fluid pass through a very fine transparent tube on which a thin beam of laser light is incident, the light transmitted and scattered by the passage of cells through the tube is collected by means of devices detection, allowing inferences to be made regarding the size and complexity of the cells. The use of fluorescent molecules makes it possible to analyze the presence of several markers simultaneously. Those known as separators or "sorters" can also purify populations of determined characteristics in different final fractions while counting the number of cells present in the sample.

The objective of this unit is:

  • Offer access to specialized equipment in flow cytometry and cell separation to INIBICA researchers and external users, providing technical expertise and training.
  • Offer technical support and advice to the research groups of the center or visiting researchers in the design of experiments, selection of markers, acquisition of samples and analysis of cell populations and discussion of the results obtained.
  • Isolation of cell populations using “Cell Sorting” technology.
  • Development of new technologies and evaluation of new equipment.
  • Training in this technology for users.
EQUIPMENT
  • Research Unit of the Puerta del Mar University Hospital:
    • Cytometer, FACSCelesta (Becton Dickinson).
    • Cell separator, FACSMelody (Becton Dickinson).
    • Cytometer, FACSCalibur (Becton Dickinson): equipped with two Lasers (Air-cooled, 15mw Argon Laser, operates at 488nm and 635nm Diode Laser). It can simultaneously measure 6 parameters, Forward Scatter, Side Scatter and 4 fluorescences.
    • Magnetic cell separator, autoMACS Pro SEPARATOR (Miltenyi Biotec).
    • Laminar flow cabinet MHD 50546/1.
  • SC-IBM:
    • Analytical Flow Cytometer Cytoflex S. Software CytoExpert Beckman Coulter.
    • Preparative Flow Cytometer Sorter MoFlo Astrios EQ Beckman Coulter in a SterilGad cabinet.
    • Heraeus Lamin Air HB2448 laminar flow hoods.
    • Fujitsu computer with Beckman Coulter's Kaluza analysis software for flow cytometry data.
    • Equipment for refrigerating samples of the Thermo A28 sorter.
APPLICATIONS

There is a wide range of flow cytometry applications, including but not limited to:

  • Multicolor analysis of cell surface and intracellular antigens (immunophenotyping).
  • Quantification of cell populations of different biological tissues.
  • Functional state of the cell: measures of viability, apoptosis, necrosis.
  • Analysis of DNA content and cell cycle.
  • Functional analysis, such as cell proliferation (CFSE), calcium flux, intracellular pH, ROS production.
  • Analysis of gene expression through the use of reporter genes.
  • Microvesicle analysis.
  • Analysis of microorganisms.
  • Separation of cell populations by flow cytometry (cell sorting).
SERVICES
  • Use (without a technician) of flow cytometry analyzers (only for internal users duly trained in the use of the equipment).
  • Use (with technician) according to the researcher's need.
  • Sorting (you can only use this service assisted by a technician).
  • Image processing and analysis. Flow cytometry data analysis.
  • Advice and support in experimental design, sample preparation methods, interpretation and presentation of data.
  • Develop / implement new flow cytometry applications / techniques that respond to user needs.
CONTACT PERSONS