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Cell Imaging, Grenoble, France

The Cellular Imaging platform allows the access to a confocal microscope (spinning disk S-M4D), a video-microscope (V-M4D), a super-resolution microscope (SMLM SR-M4D) and a flow-cytometer (VYB). The instruments are optimised to study living cells, and fluorescent protein expression. Details are available on the ISBG website.

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User Guide

Spinning disk confocal microscope 

The spinning disk confocal microscope (S-M4D; Olympus and GATACA) has been upgraded in May 2022. It is configured with a Nipkow disk (Yogokawa CSU-X1) and 6 solid state lasers for confocal imaging in real time on living cells (very low phototoxicity). Acquisition is performed using an EMCCD camera (Andor iXon ultra ; MetaMorph). A photoconversion/photoactivation module is available for FRET and FRAP (Ilas2).

Time-lapse video-microscope 

The time-lapse video-microscope (V-M4D, Olympus), renewed in 2019, is composed of an inverted Olympus IX83 fully motorized microscope equipped with a blind, temperature controlled, incubation chamber (Digital Pixel) for live imaging. Acquisition is achieved with a sCMOS camera (Hamamatsu Orca Flash4; CellSens).

Multi-channel flow-cytometer 

The multi-channel flow-cytometer (VYB, Miltenyi biotech) is equipped with three lasers and 8 fluorescence channels. It is optimised for fluorescent proteins, and covers all the emission wavelengths used with microscopy. It is automatable (multiwell plate, immuno-staining, dilutions), and compatible with magnetic columns Miltenyi (isolation of rare events).

Analysis

Cytometry and microscopy analysis software are available for users in a dedicated room. Three workstations are installed with Volocity and Imaris for microscopy data visualization and analysis; or MacsQuant and CellQuest for flow cytometry.

Super-resolution PALM/STORM microscope.

The system SAFe360 from Abbelight offers a 3D multicolor imaging (including "spectral demixing") of subcellular structures as well as a single particle tracking capabilities. This new setup is highly automated for a better user experience. It is equipped with a laser combiner hosting 6 high-power lasers, a module for homogeneous sample excitation in EPI/ HiLo and TIRF modes, a high numerical aperture x100 oil immersion objective, and two sensitive and rapid sCMOS cameras allowing the acquisition rates of more than 100 frames per second. 
ISBG M4D platform website: https://www.isbg.fr/cellular-analysis/cellular-imaging/ 
Abbelight website: https://www.abbelight.com/solutions/abbelight-instruments/#SAFe360