Cytometry Platform

The Flow Cytometry and Experimental Irradiation Research Support Platform makes available to researchers, both belonging to and external to ibs.GRANADA, the equipment and scientific-technical advice necessary to carry out the dispersion analysis. of light and different fluorescence in cells, microorganisms or subcellular units such as exosomes, among others. Furthermore, separation is possible (sorting) in populations or homogeneous groups of interest. All of this aimed at facilitating and developing quality research.

This platform is characterized by having, among its multiple Service Portfolios, a portfolio for the Immunophenotyping of lymphomas and leukemias in experimental animals, which is currently a national reference.

Located on the 1st Floor, Old Management Building, San Cecilio Hospital, Avd. Madrid sn de Granada.

There are two types of service provision:

1. Use of the equipment, self-service or with the support of the Responsible Technician. Due to the complexity of using this equipment, the researcher must provide certification of knowledge of it. The cost of using the equipment is specified in the document "Rates".
2. Performance of Specific Techniques by the Responsible Technicians. The cost of these techniques depends on their complexity and the number of samples to be analyzed, and must be individually budgeted.

• • CF_01 - Quantification of cytokines, cell signaling proteins, soluble and immunoglobulins by Flow Cytometry. ABC method
  • CF_02 - Immunophenotyping of lymphomas and leukemias in experimental animals by flow cytometry
  • CF_03 - Characterization and isolation of Cancer Stem Cells by flow cytometry
  • CF_04 - Qualitative and quantitative determination of DNA damage by flow cytometry
  • CF_05 - Characterization of lymphocyte populations and subpopulations by flow cytometry
  • CF-06 Studies of cell apoptosis by flow cytometry (Annexin V)
  • CF_07- Analysis of the cell cycle by determining the cell phase by Flow Cytometry
  • CF-08 Evaluation of the expression of kappa/lambda light chains in B-cell lymphomas by flow cytometry
  • CF_09- Characterization and isolation of Cancer Stem Cells based on their Aldh-1 and AldeRed activity by Flow Cytometry
  • CF_10- Study of the production of reactive species in leukocytes by flow cytometry
  • CF_11- Study of Cell Proliferation by Labeling with CellTrace CFSE by Flow Cytometry
  • CF_12- Study of the variation of the mitochondrial membrane potential (Δψm) by Flow Cytometry (JC-1)

BD FACS Melody Flow Cytometer This equipment has 3 laser lines:

• Violet Laser (405 nm): Equipped with 3 fluorescence detectors
• Blue Laser (488 nm): equipped with 4 fluorescence detectors and 2 light scattering detectors.
• Red laser (638nm): It has 2 fluorescence detectors

It has a cell separation system that allows obtaining up to 4 homogeneous pure populations in different supports (Cytometry tubes, 1.5 mL Eppendorf or 15 ML Falcon). In addition, it is equipped with the cloning/single cell unit that allows the separation of one or more cells per well in multiwell plates.

The equipment has a workstation equipped with BD FACS Chorus Software for equipment control and data analysis.

Upgraded BD FACS ARIA III Flow Cytometer: with 4 laser lines and 16 fluorescence detectors (18 simultaneous parameters):

• Violet laser (405 nm), equipped with 7 fluorescence detectors
• Blue laser (488nm), allows to detect 2 fluorescence and the morphometric parameters FSC/SSC
• Yellow/green laser (561 nm), has 4 fluorescence detectors
• Red laser (633nm), equipped with 3 fluorescence detectors

With a cell separation system (up to 4 populations) and the Single Cell/Cloning unit (automatic cell deposition unit) that allows the deposition of one or more cells in multiwell plates, slides and tubes from 0,5 to 15 mL (Index system Sort).

All with acquisition and analysis software Diva 8

Cytek Northern Lights Spectral Cytometer: This equipment consists of 3 laser lines and a total of 38 detectors:

• Violet laser (405 nm) with 16 detectors for this laser.
• blue laser (488 nm) with 14 detectors:
• red laser (638nm) with 8 detectors.

This allows detecting up to 38 parameters simultaneously.

The detectors are high quantum efficiency avalanche photodiodes (APDs).

A large number of fluorochromes can be used at the same time, since it covers the spectrum from 420 to 829 nm.

It has three light scatter channels, one forward scatter (Forward Scatter, FSC) and two side-scatter (Side Scatter, SSC):

• FSC: high performance detector with a 488 nm bandpass filter.
• SSC Violet: High performance detector with a 405 nm bandpass filter.
• SSC Blue: High performance detector with a 488 nm bandpass filter.

The team has a workstation equipped with the Software SpectroFlo, for data acquisition and analysis.

YXLON MaxiShot E200 Irradiator: equipment created specifically for cellular radiation in experimental processes.

It allows irradiation to be carried out at low doses (from 1Gy) and high doses, so it can be useful in a wide variety of tests such as:

· The analysis of cellular and organic response after being exposed to this type of stress.

· Survival kinetic studies after different doses of radiation

· Determination of the effectiveness of combined therapy (radio and chemotherapy) as treatment

· Studies of DNA repair capacity.

· Basic studies of cellular and molecular radiosensitivity