Flow Cytometry and Cell Culture Section

Presentation

The cell culture and flow cytometry section of the Central Medicine Research Unit (UCIM), is a laboratory dedicated to the development of a set of techniques that allow the cataloging, maintenance and study of cells “in vitro” as well as cryopreservation of cell lines. The section comprises 3 work areas:

• Cell cultures
• Flow cytometry
• Cryopreservation

Flow cytometry is a technique that allows us to measure and obtain information on the optical characteristics of cells or particles in suspension when subjected to a laser light source.

This measurement is made on each individual particle based on forward scatter, related to cell size, 90-degree lateral scatter, related to internal structure and cytoplasmic complexity, and emission fluorescence of particles illuminated by laser light.

Flow cytometry allows us to study the phenotypic characteristics of cell populations, the immunophenotype, using fluorescent monoclonal antibodies, and the functional characteristics using the different available fluorescent probes.

Flow cytometry is a fast system, being able to analyze thousands of cells/minute, and multiparametric, allowing the analysis of various cellular parameters simultaneously.

The purpose of the section is to provide scientific and technical advice to different users, both internal and external, for which different flow cytometers, analyzers, separators and an image cytometer system, microscopes, flow cabinets, CO2 incubators are available in different O2 percentages and a room for cryopreservation of samples.

Process

The user must contact the service by phone or email, to be informed of budgets and facilities.

Once the budget has been accepted, the user will take their samples in hermetic containers that prevent contamination of personnel and the environment in the event of a spill.

In case of requesting sample analysis by flow cytometry, contact those responsible to be informed of the sample staining protocols.

If the user works with cell lines from our sample bank in a state of cryopreservation, they must follow the different expansion and thawing protocols.

Services

Cell Cultures

• Analysis of cell culture quality (mycoplasma evaluation, cell culture treatment).
• Maintenance of primary cell culture lines.
• Culturing under different O₂ concentrations.
• Listing, storage, and cryopreservation of various types of cell lines.

Flow Cytometry

• Studies of cell viability and proliferation: cell death, apoptosis, IC50 determination.
• Advice on the preparation of cytometric protocols. Design of multicolor panels. Sample processing.
• Immunophenotyping of human blood cell populations.
• Immunophenotyping of blood cell populations from various animal species: mouse, rat, pig, rabbit.
• Study of cell populations in biopsy samples.
• Study of physiological processes at the cellular level: Cell cycle, Apoptosis, Mitochondrial markers, Ca++ levels, DNA damage analysis, Oxidative stress.
• Study of intracellular cytokine levels.
• Cell sorting in cell culture samples as well as patient samples. Sorting of cells transfected with fluorescent vectors. Sorting of leukocyte populations of interest.
• Imaging flow cytometry: morphometric studies by image, detection of colocalization of fluorescent probes.
• Physical cytometry: analysis and sorting of spheroids and organoids [50–500 μm].
• Analysis of cytometry data generated by the service using specialized software.
• Basic cytometry course for users.

Equipment

• Single pan analytical and precision balance
• Analogue bain-marie
• 3D bioprinter
• High microbiological safety cabinet
• Type IIA Biological Safety Laminar Flow Cabinet
• Laminar flow cabinet
• Refrigerated centrifuge with rotors – Eppendorf
• Refrigerated centrifuge with rotors – Heraeus
• Analyzer Flow Cytometer – BD FACSVERSE
• Analyzer Flow Cytometer – BD LSR Fortessa X-20
• Separator Flow Cytometer-BD FACS AriaIII
• Amnis ImageStream imaging flow cytometer. Luminex
• W8 physical cytometer. Separator of spheroids and organoids. Cell Dynamics
• Magnetic separation system. Miltenyi
• Refrigerated centrifuge with rotors – Gyrozen
• Automatic cell counter
• Spectrofluorimeter
• Low Volume Spectrophotometer
• Spectrophotometer
• Incubator 5% CO₂ 21%O 2 – Heracell 150i
• Incubator 5% CO₂ 21%O2 – SANYO MCO18AIC
• Incubator 5% CO₂ 21%O2 – SANYO MCO19AIC
• Incubator 5% CO2 y 21%O2 – Thermo Fischer FORMA STERICYCLE
• 5% CO₂ 21%O 2 incubator for organoids
• Incubator hipoxia 5% CO2 0-21% O2 – Thermo Fischer FORMA STERICYCLE i160
• Refrigerated microcentrifuge
• Inverted microscope
• N2 tank
• Thermal cycler
• Ultra centrifuge