CYTOMETRY AND SEPARATION
Flow 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 based on their morphological characteristics and the presence of biomarkers, as well as in protein engineering. In flow cytometers, cells suspended in a fluid pass through a very fine transparent tube onto which a narrow laser beam is directed; the light transmitted and scattered as the cells pass through the tube is detected by sensing devices, allowing inferences to be made regarding the size and complexity of the cells. The use of fluorescent molecules allows for the simultaneous analysis of various markers. 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:
- To provide INiBICA researchers and external users with access to specialized flow cytometry and cell separation equipment, while offering technical expertise and training.
- Provide technical support and guidance to the center’s research groups or visiting researchers in the design of experiments, selection of markers, collection of samples, and analysis of cell populations, as well as in the 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 (a 15 mW air-cooled argon laser operating at 488 nm and a 635 nm diode laser). It can simultaneously measure 6 parameters: forward scatter, side scatter, and 4 fluorescence channels.
- Magnetic cell separator, autoMACS Pro SEPARATOR (Miltenyi Biotec).
- MHD 50546/1 Laminar Flow Cabinet.
- Cytoflex S Analytical Flow Cytometer. Beckman Coulter CytoExpert software.
- Beckman Coulter MoFlo Astrios EQ preparative flow cytometer in a SterilGad enclosure.
- Heraeus Lamin Air HB2448 Laminar Flow Hoods.
- Team Fujitsu computer equipped with Beckman Coulter's Kaluza analysis software for flow cytometry data.
- Team to cool samples from 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 in different biological tissues.
- Cell functional status: measures of viability, apoptosis, and necrosis.
- Analysis of DNA content and the cell cycle.
- Functional analysis, such as cell proliferation (CFSE), calcium flux, intracellular pH, and ROS production.
- Gene expression analysis using marker 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 a technician) as needed by the researcher.
- Sorting (this service can only be performed with the assistance of a technician).
- Image processing and analysis. Flow cytometry data analysis.
- Advice and support on experimental design, sample preparation methods, and data interpretation and presentation.
- Develop and implement new flow cytometry applications and techniques that meet user needs.
RESERVATIONS
If you have any trouble downloading the file mentioned above, you can access it via the following link.
CONTACT PERSONS
- SC-IBM UCA: dg.recursoscientificos@uca.es
- U.I.H.U.P.M: laura.jaramillo@inibica.eu