Service / Technology Instance

About Cell Imaging, Grenoble, France

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

Instruments Available:

Our flow-cytometer (VYB, Miltenyi biotec) is installed within a L2 facility (room 547C), and equipped with three diode lasers and 8 fluorescence channels. It is optimized for fluorescent proteins detection, and covers all the emission wavelengths used with the epi-fluorescence and confocal microscopes of the cell imaging facility. It is fully programmable (automated labelling and sample manipulation) and is compatible with multi-well plate. A dedicated workstation for analysis is also available for users.

The confocal spinning disk microscope (S-M4D) is optimized to study living cells. It is installed since January 2013 in room 547B. It is composed of an epifluorescence Olympus IX81 microscope equipped with plan-apo 20x dry, 40x dry and oil, and 60x and 100x immersion objectives (with wide numerical apertures) and with differential interference contrast (Nomarski). This inverted and fully motorized microscope is equipped with an incubation chamber and with a CO2 controled microenvironmental chamber for live imaging. IQ2/Imaris (Andor/Bitplane) is used for controlling the microscope and for treatment of the acquisitions. Volocity (Perkin Elmer) is also available to users on a specific workstation.

The video-microscope (V-M4D) is installed in room 547D. It is composed of an epifluorescence Olympus IX81 microscope equipped with plan-apo 20x (dry), 40x, and 60x immersion objectives (with wide numerical apertures) and with differential interference contrast (Nomarski). This inverted and fully motorized microscope has been installed within an incubation chamber (Solent) set at 37°C for live imaging. Volocity (Perkin-Elmer) is used for controlling the microscope and for treatment of the acquisitions.

User Guide

Spinning disk confocal microscope 

The spinning disk confocal microscope (S-M4D; Olympus and Andor) is configured with a Nipkow wheel (Yogokawa CSU-X1) and 6 solid state lasers for confocal imaging in real time on living cells (very low phototoxicity). Images acquisition are done simultaneously with 2 EMCCD cameras (Andor iXon ultra). A photoconversion/photoactivation module is available for FRET and FRAP.

Time-lapse video-microscope 

The time-lapse video-microscope (V-M4D, Olympus and Perkin Elmer) is, as for the confocal composed of an inverted Olympus IX81 fully motorised microscope equipped with a piezo stage, and an incubation chamber set at 37°C for live imaging. Acquisition is achieved with a sCMOS camera (Hamamtsu Orca Flash4; Volocity).

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 (multiwells 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 visualisation and analysis; or MacsQuant and CellQuest for flow cytometry.

Super-resolution PALM/STORM microscope.

This is a noncommercial in-house built setup based on a microscope (Olympus IX81), 2 EMCCD cameras (Photometrics) and 5 lasers (405 nm, 488 nm, 532 nm, 561 nm, 643 nm) controlled by an acousto-optic filter (AOTF). The EMCCD cameras are controlled through the Metamorph software, the lasers are controlled through LabVIEW. More details on the IBS website. This instrument is only open to collaborations (no funding)

Instruct Centre

Instruct Centre FR2

IBS-ISBG

71 avenue des Martyrs

Grenoble

France

www.ibs.fr & www.isbg.fr

Cell Imaging, Grenoble, France

Contacts:

Françoise Lacroix
Françoise Lacroix
Institut de Biologie Structurale
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Jean-Philippe Kleman
Jean-Philippe Kleman
Institut de Biologie Structurale
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