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Integrated Structural Biology Grenoble
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Contact person(s) related to this article / INDORATO Rose-Laure / KLEMAN Jean-Philippe / LACROIX Françoise

Cellular Imaging

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M4D 002©CEA/D. Morel

Presentation

The Cellular Imaging platform grants the access to a confocal microscope (spinning disk S-M4D), a video-microscope (V-M4D), and a flow-cytometer (VYB). The instruments are optimized to study living cells.

1 - The confocal microscope (S-M4D; Olympus and Andor) is configurated 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.

2 - The video-microscope (V-M4D, Olympus and Perkin Elmer) is, as for the confocal composed of an inverted Olympus IX81 fully motorized 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)

3 - The flow-cytometer (VYB, Miltenyi biotech) is equipped with three lasers and 8 fluorescence channels. It is optimized 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).

4 - 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.

5 - 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 web site.

Key words

Microscopy; DIC; Epifluorescence; 4D, Flow-cytometry ; Spinning-disk confocal

Dedicated Staff

The technical and scientific expertise, and the practical management of the platform are under the responsibility of RL Revel-Goyet, F. Lacroix and JP Kleman.

Accessibility

Authorized users can access freely the equipment of the platform (microscope and post-treatment dedicated computer), after they have validated the internal training session, and accepted the conditions of use.
For more complex projects or if a specific expertise is requested (collaboration), we can provide a technical assistance for some or all of the experiments planned (observation and post-treatment).

How to make a request?

Requests are made through the online form or by direct contact.

Contact and localisation

The platform is located room 547 in the IBS.

Samples

The microscope is dedicated to fixed or living Eukaryote and Procaryote cells observation (level 2). Security training is mandatory to access to the room. A tissue culture room is available for customers (L2).

Results

In general, the acquisition data and the analyzed data are stored for 1 year. A spare copy is automatically generated, but the platform cannot guarantee the integrity of the data stored in case of dysfunction. The user is responsible for the final storage of his own datasets. After one year, the data can be erased by the platform supervisors without notification.

Cost

Users are charged for the operation costs calculated per hour . Contact us for further info.

Follow-up/Valuation

The platform can help users who need technical advises (preparation of samples) or during treatment and analysis of the data.

Users should cite the platform in their publications :
« We thank Rose-Laure Revel-Goyet, Françoise Lacroix and Jean-Philippe Kleman (Institut de Biologie Structurale, Grenoble) for the support and access to the Cell imaging Platform. ») et remercier FRISBI et GRAL pour le support de l’UMS :"This work used the platforms of the Grenoble Instruct-ERIC Center (ISBG : UMS 3518 CNRS-CEA-UGA-EMBL) with support from FRISBI (ANR-10-INBS-05-02) and GRAL (ANR-10-LABX-49-01) within the Grenoble Partnership for Structural Biology (PSB)."
For collaborative work, supervisors of the platform must co-sign papers.

web site

Intranet web site (french only) is accessible from the IBS network.