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Flow cytometer CANTOII^© (Beckton Dickinson, Fluorescent Automated Cell Sorting)

Flow cytometry is a technique that enables the simultaneous measurement and analysis of numerous physical parameters (relative size, granularity and fluorescence) of suspended particles between 0.1 and 150 μm in size. These characteristics are measured using an optical system coupled to an electronic system, which records the scattering of incident light and the emission of fluorescence by particles exposed to a laser excitation source.

Flow cytometry can be used for a variety of applications: phenotypic characterisation, measurement of intracellular cytokine production, modulation of membrane antigen expression after cell activation or transfection, cell cycle analysis, analysis of intracellular calcium concentration, etc.

The CANTOII cytometer, obtained with financial support from the Ile de France region (DIM Maladies Infectieuses) and available at BIPAR JRU, has 3 lasers (405nm, 488nm, 633nm) and can detect up to 10 different parameters simultaneously, including 8 fluorescence parameters. It is also equipped with a High Throuput Sampler (HTS).

It is open to researchers from the EnvA and ANSES research laboratories, on request to Delphine Le Roux.

Wide-field Laser scanning confocal microscope DMi8 (Leica)

The JRU is equipped with a DMi8 wide-field confocal laser scanning microscope (CLSM).

This is an inverted microscope equipped with:

- objectives 10x (ON 0.40), 20x (ON 0.7) immersion, 40x (ON 1.30) oil, 63x (ON 1.40) oil, 100x (ON 1.40) oil

- motorised board

- tandem scanner comprising a conventional 22mm linear FOV scanner + and a 13mm linear FOV resonant scanner (8000 Hz; 29 images/second at 512*512)

- Diode lasers 405, 488, 552, 638 nm

- 4 sensors: 1 PMT et 2 HyD (ultra-sensitive for fluorescent, 1 PMT for transmission)

- Sola SMII LED fluorescent source and long-pass filters for observing fluorescent light through eyepieces

- chamber with temperature (+5°C to +50°C) and CO2 pour live imaging

Confocal microscope (^©G. Karadjian, BIPAR JRU)