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Cytometry

The platform allows the phenotyping of cells through the detection and identification of membrane and intracellular molecules. In order to purify a cell subpopulation, we also perform high throughput cell sorting, based on multi-parametric cell analysis. These subpopulations can then be studied for different parameters (transcriptome, proteome, in vivo transfer...).

RESTORE cytometry platform is a partner of the Toulouse Imaging Network Platform (TRI). It is certified ISO 9001 version 2015. The platform has a cell sorter, two analyzers and an image cytometer and offers a strong expertise on cells coming from solid tissues (adipose tissue, muscles, gums…). All the equipment is integrated in a L2 containment area.

The use of the equipment is open to research teams from public and private institutions, subject to acceptance of the projects by the manager.

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expertise

Flow cytometry is a technique for the individual, quantitative and qualitative characterization of particles suspended in a flow.

Fortessa X20 (BD biosciences)

The Fortessa X20 is equipped with 5 lasers: 488nm blue, 405nm violet, 355nm UV, 640nm red, 561nm yellow-green, capable of reading 19 parameters. The Fortessa on the platform is also equipped with a 96-well plate changer. The platform is equipped with DIVA, Flowlogic and Kaluza analysis softwares, which can be used on site under reservation conditions.

MACSQuant 10 (Miltenyi)

The MACSQuant is equipped with 3 lasers: 405nm, 488nm and 640nm, which can read 10 parameters. This cytometer is also equipped with a plate changer, a marking and automatic dilution module. It allows to standardize experiments.

The analyzers are freely available to users who have undergone mandatory training by the platform staff.

Cell sorting by flow cytometry allows the isolation and recovery of one or more cell subpopulations according to size, granulority and fluorescence criteria. For example, we are able to sort cells representing 0.1% of the vascular stromal fraction of adipose tissue.

ARIA FUSION
ARIA FUSION

ARIA FUSION

The sorter present on the plateforme is an ARIA Fusion (BD Biosciences) under sterile environment equipped with 4 lasers: 561nm yellow-green, 407nm violet, 633nm red, 488nm blue, able to read 18 parameters.

The sorter present on the tray is equipped with the following accessories:

  • 4 way sorting
  • sorting in tubes of 1.5ml, 2ml, 5ml, 15ml
  • 70µm, 85µm, 100µm nozzle
  • refrigeration module at the sorting’s inlet and outlet.

Sorting is carried out by the platform staff as a service at the request of the research teams and after acceptance by the platform manager.

Flow cytometry imaging is a technology that allows the visual power of microscopy and the statistical rigor of flow cytometry to be combined in a single instrument.

Image Stream X
Image Stream X

Image Stream X

The specific applications of the Image Stream X are multiple, we can mention in particular :

  • morphology analysis,
  • internalization,
  • cell signaling and molecular translocation,
  • co-localization, cell interactions, cell cycle, apoptosis,
  • spot counting,
  • autophagy and cell death.

The platform has an imaging flow cytometer: Image Stream X Mark II (Merck-Luminex). This apparatus is equipped with 3 objectives (20x, 40x and 60x), 3 lasers (405nm, 488nm and 642nm) and a camera and can therefore read 6 parameters simultaneously.

The image files consisting of tens of thousands of cell images can be routinely analyzed using the IDEAS software: analysis of up to 500 morphometric and photometric parameters for each cell image using patented algorithms. The software is present on the platform and can be used under reservation conditions.

Acquisitions are carried out by the platform staff as a “service” at the request of the research teams and after acceptance by the platform manager. Analyses can be carried out with the help or under the supervision of the platform personnel, in the form of a service or collaboration.

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Marie-Laure Renoud 2

Marie-Laure Renoud

Engineer

The platform's engineer evaluates the user's request and proposes the most suitable solution for his needs:

  • providing advice in the development of multi-color panels,

  • providing training to users,

  • assisting users in the analysis and interpretation of their results,

  • assisting users in the implementation of research and development projects, in the form of collaboration.

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Equipements

Pour les utilisateurs réguliers : Réservation directe