Flow Cytometry and Sorter
Mission
The Flow Cytometry and Sorter Unit is a Research Support Service (SAI) of the IiSGM. The Unit’s mission is to provide flow cytometry support to researchers, both from the IiSGM and external researchers or private companies. The Unit offers scientific and technical support to researchers, ranging from advice on experimental design and data interpretation to the use of computer software for multiparametric analysis. This enables the discovery, identification and monitoring of cellular markers, while also facilitating the separation of the different cellular populations identified.
Description
Flow cytometry is an analytical/quantitative technique that enables multiple characteristics of cells or particles of interest contained in a fluid to be measured through labelling with antibodies or fluorescent probes. Interaction with one or more lasers results in light scattering and fluorescence, which is translated into detailed information about the structural and functional characteristics of the cells or particles under study. The application of the Cell Sorter allows cell populations to be separated under sterile conditions according to the characteristics analysed by flow cytometry.
This technique allows multiple applications to be carried out simultaneously in a reduced sample volume. It is important to emphasise that this technique performs measurements particle by particle, beyond descriptive population measurements. The identification of cell markers is applicable to the research, diagnosis, monitoring and treatment of a wide range of diseases, making Flow Cytometry and Cell Sorting a technology involved in translational research.
The Flow Cytometry Unit has been certified since 2016 in accordance with the guidelines of the Quality Management System (ISO 9001:2008), and since 2019 it has complied with the requirements of the updated standard (ISO 9001:2015).
Services offered
Flow Cytometry Analyser
- Phenotyping of different lymphocyte markers on the surface or intracellularly.
- Analysis of cell viability through staining with propidium iodide, annexin V or 7-AAD.
- Cell cycle analysis using propidium iodide.
- Analysis of transfection efficiency.
- Quantification of soluble cytokines in plasma, serum or culture supernatants using a multiplex assay (up to 11 cytokines in 25ul of sample).
- Analysis of intracellular cytokines.
- Analysis of intracellular calcium movement using Fluo-4 staining.
- Analysis of apoptosis activation using various markers. E.g. caspase-3.
- Analysis of DNA fragmentation using the TUNEL technique.
- Detection of microparticles and exosomes in culture supernatants, plasma and serum.
Flow Cytometry Cell Sorter
- Cloning.
- Separation of cell populations of interest.
- Separation of live cells for cell culture.
- Separation of fixed cells for subsequent genetic analysis.
Support for Researchers
- Advice and optimisation of study protocols or experimental design.
- Training on demand and assessment of researchers' needs.
- Personalised advice on experimental approaches, fluorochromes and available markers.
- Standardised procedures for the best possible processing of samples.
- Analysis using various software applications.
Rates available at www.iisgm.com (Research Support Services).
Technology
Flow cytometer 10 colours, 12 parameters. Model Gallios (Beckman Coulter)
- Detection of 10 colours, 12 parameters. Three lasers (blue-488nm, red-638nm and violet-405nm), computer-controlled, with a rapid start-up and stabilisation system.
- Next-generation photomultiplier tubes, with high sensitivity in the red range and excellent linearity throughout the range.
- Possibility of analysing rare events due to both the high acquisition speed and the possibility of acquiring large sample volumes.
Flow cytometer 14 colours, 16 parameters. Model MAQSquant 16 (MiltenyiBiotec)
Detection of 14 colours, 16 parameters. Three lasers: blue-488nm (6 detectors), red-640nm (3 detectors) and violet-405nm (5 detectors).
- Possibility of selecting the FSC with the violet laser to increase resolution when analysing small particles.
- Sample acquisition in several containers: tubes, Eppendorf tubes and 96-well plates (with refrigeration option).
- Acquisition of small volumes.
Flow cytometer sorter, model MoFlo Astrios (Beckman Coulter)
- Integration into a type II biological safety cabinet.
- Sample aerosol evacuation and agitation system.
- Controlled sample temperature (cooling or heating).
- Possibility of up to 6 solid-state lasers (640nm, 592 nm, 561 nm, 532 nm, 488nm, 405 nm) and 7 phinoles. It currently has 2 lasers (488nm and 640nm).
- Depending on the application, the acquisition speed can be up to 100,000 cells/sec; automatic recognition by photomultipliers (PMTs) and PODs, enabling different configurations according to the objective.
- Various collection systems: multi-well plates, propylene tubes, etc.
Fluorescence microscope model Eclipse 50i with optical system from Nikon CFI60
Installation
The SAI Flow Cytometry and Sorter is located in the Experimental Medicine and Surgery building
in the IiSGM prefabricated pavilion. The room has the appropriate electrical grid, air conditioning and temperature conditions to meet the quality requirements of the services offered.
Contact
Dra. Laura Díaz Muñoz
Head of the Unit
E-mail: lauradiaz(ELIMINAR)@gmail.com
Tel: 91 47 92 58
Experimental Medicine and Surgery Pavilion (1st floor)
Hospital General Universitario Gregorio Marañón
C/ Doctor Esquerdo, 46; Madrid 28007
Website with the booking procedure: umce.hggm.es