CYTOMETRY AND SEPARATION
Flow cytometry 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 to count and classify cells according to their morphological characteristics, the presence of biomarkers, and in protein engineering. In flow cytometers, cells suspended in a fluid pass through a very fine transparent tube onto which a thin laser beam is directed. The light transmitted and scattered by the passage of the cells through the tube is collected by detection devices, allowing inferences to be made about the size and complexity of the cells. The use of fluorescent molecules allows the presence of several markers to be analyzed simultaneously. Devices known as sorters can also purify populations with specific characteristics into different final fractions while counting the number of cells present in the sample.
The objective of this unit is:
- Provide access to specialized flow cytometry and cell separation equipment to INiBICA researchers and external users, offering technical expertise and training.
- Provide technical support and advice to the center's research groups or visiting researchers in the design of experiments, selection of markers, acquisition of samples, 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
- U.I.H.U.P.M:
- Flow cytometer, FACSCelesta (Becton Dickinson).
- Cell separator, FACSMelody (Becton Dickinson).
- Flow cytometer, FACSCalibur (Becton Dickinson): equipped with two lasers (air-cooled 15 mW argon laser operating at 488 nm and 635 nm diode laser). It can simultaneously measure six parameters: forward scatter, side scatter, and four fluorescences.
- Magnetic cell separator, autoMACS Pro SEPARATOR (Miltenyi Biotec).
- MHD 50546/1 laminar flow cabinet.
- Cytoflex S Analytical Flow Cytometer. CytoExpert Software Beckman Coulter.
- Beckman Coulter MoFlo Astrios EQ preparative flow cytometer in SterilGad cabinet.
- Heraeus Lamin Air HB2448 laminar flow hoods.
- Fujitsu Team with Beckman Coulter's Kaluza analysis software for flow cytometry data.
- Team cooling samples from the Thermo A28 sorter.
APPLICATIONS
There is a wide range of flow cytometry applications, including, among others:
- Multicolor analysis of cell surface and intracellular antigens (immunophenotyping).
- Quantification of cell populations from different biological tissues.
- Functional state of the cell: measurements of viability, apoptosis, necrosis.
- DNA content and cell cycle analysis.
- Functional analysis, such as cell proliferation (CFSE), calcium flux, intracellular pH, ROS production.
- Gene expression analysis using reporter genes.
- Microvesicle analysis.
- Analysis of microorganisms.
- Separation of cell populations using flow cytometry (cell sorting).
SERVICES
- Use (without a technician) of flow cytometry analyzers (only for internal users who have been properly trained in the use of the Team).
- Use (with technician) according to the researcher's needs.
- Sorting (this service can only be used with the assistance of a technician).
- Image processing and analysis. Flow cytometry data analysis.
- Advice and support in experimental design, sample preparation methods, data interpretation, and presentation.
- Develop/implement new flow cytometry applications/techniques that meet user needs.
RESERVATIONS
If you have any problems downloading the above file, you can access it via the following link link.
CONTACT PERSONS
- SC-IBM UCA: dg.recursoscientificos@uca.es
- U.I.H.U.P.M: laura.jaramillo@inibica.eu