Cytometry and Cell Sorting Units
Flow cytometry is a cell analysis technique that involves measuring the light scattering and fluorescence characteristics of cells as they pass through a beam of light. Flow cytometry is used in the counting and classification of cells according to their morphological characteristics and presence of biomarkers, as well as in protein engineering. In flow cytometers, cells suspended in a fluid pass through a very thin transparent tube that is hit by a thin beam of laser light. The light transmitted and dispersed by the passage of the cells through the tube is collected by detection devices, allowing to make inferences on size and complexity of the cells. The use of fluorescent molecules allows to analyse the presence of several markers simultaneously. Sorters can also purify populations with specific characteristics into different final fractions while counting the number of cells present in the sample.This unit aims to:
- Provide access to equipment specialized in flow cytometry and cell sorting to INiBICA's researchers and external users, while providing technical expertise and training.
- Provide technical support and advice to INiBICA's research groups or visiting researchers in the fields of design of experiments, selection of markers, acquisition of samples, analysis of cell populations and discussion of the results obtained.
- Provide isolation of cell populations through "Cell Sorting" technology.
- Develop new technologies and assess new equipment.
- Provide training on this technology to users.
- Cytometer, FACSCalibur (Becton Dickinson): equipped with two lasers (air-cooled laser Argon 15mw operating at 488nm and diode laser operating at 635nm). 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
- Analytical flow cytometer: Cytoflex S.Software CytoExpert Beckman Coulter.
- Preparative flow cytometer sorter: MoFlo Astrios EQ Beckman Coulter in SterilGard cabinet.
- Laminar flow hoods: Heraeus Lamin Air HB2448.
- Fujitsu computer equipment with Beckman Coulter's Kaluza analysis software for flow cytometry data.
- Sample cooler equipment for Thermo A28 sorter.
There is a wide range of flow cytometry applications, including but not limited to:
- Multicoloured analysis of cell surface and intracellular antigens (immunophenotyping).
- Quantification of cell populations of different biological tissues.
- Functional status of cells: measures of viability, apoptosis, necrosis.
- DNA content and cell cycle analysis.
- Functional analysis, such as cell proliferation (CFSE), calcium flow, intracellular pH, ROS production.
- Gene expression analysis by means of reporter genes.
- Microvesicle analysis.
- Analysis of microorganisms.
- Sorting of cell populations by flow cytometry (cell sorting).
- Use (without technician) of flow cytometry analysers (only for internal users properly trained in the use of the equipment).
- Use (with a technician) according to the needs of the researcher.
- Sorting (this service can only be used with the assistance of a technician).
- Image processing and analysis. Analysis of flow cytometry data.
- Advice and support in experimental design, sample preparation methods, data interpretation and presentation.
- Development / implementation of new applications / flow cytometry techniques that meet user's needs.
- SC-IBM: firstname.lastname@example.org
- U.I.HUPM: email@example.com