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MitoVasc : physiopathologie cardiovasculaire et mitochondriale


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    Microscopy and Image Analysis platform

    Microscopy and Image Analysis platform

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    Persons in charge :

    Daniel Henrion, Céline Fassot and Laurent Loufrani

    Materials : 

    • Spinning-disk (Nipkow disk) confocal microscopes (Resp.: Daniel Henrion)

    The laboratory is equipped with a Spinning-disk confocal microscopes which achieve frame rates of over 50 per second a feature necessary for dynamic observations such as live cell Imaging or live tissues scanning.



    The system (solamere Technology, Salt Lake City) is mounted on an inverted NIKON TE2000 microscope (Acroplan objective 10x and 20x, Plan Fluor 40x and water immersion 63x Nikon plan Apo).
    - Camera: CoolSNAP HQ2
    - Software: Metamorph, v7.5
    - LASERS : 491 nm 50mW, 561 nm 50mW, 408 nm 44mW and 638 nm 28mW. 
    The microscope is equipped with an organ bath allowing scanning the arterial wall in pressurized and perfused isolated arteries.

    •  Real-time confocal imaging (Resp.: Nathalie C. Guérineau)

    The laboratory is equipped with two confocal microscopes allowing imaging of excitable tissues (acute living slices and cell cultures).

    1- Fast laser scanning confocal microscope (Noran confocal technology, Odyssey XL, argon-krypton laser, laser lines: 488 and 568 nm) allowing real-time imaging of intracellular calcium transients (up to 480 images/s) mounted on an upright Zeiss Axioskop FS.


    2- Fast laser scanning confocal microscope (LSM 5-Live) equipped with 3 laser lines (405, 488 and 532 nm), mounted on an upright Zeiss Axio Examiner. It allows imaging of fast fluorescent cell signals (>60 frames/s, 512x512 pixels, 2 colors). Fluorescent signals can also be visualized by epifluorescence (excitation wavelengths 365, 475 and 545 nm).



    Each of these two setups also allows electrophysiological recordings of electrical signals (single patch-clamp recordings: EPC-9 HEKA Instruments, or multiple patch-clamp recordings: EPC-10 quadro HEKA Instruments).

    •  Histo-morphometry and epifluorescence analysis (Resp.: Laurent Loufrani)

    The laboratory is equipped for histo-morphometry with an Olympus IMT 100 inverted microscopes for the histomorphometric analysis. 


    The microscope is equipped with : 

    - SONY DXC390 P tri CCD 1/3” for image analysis. 
    - Olympus objectives : 5x, 10x, 20x and 40x.
    - MuTech IV460 image acquisition car and Histolab software. 
    - Histomorphometric measurements are achieved using the Archimed Pro data base and software (Microvision, France)

     

     

     

     

     

     

     

    The laboratory is equipped for epifluorescence analysis with a Nikon E600 inverted microscopes for epifluorescence analysis.


    The microscope is equipped with:
    objectives : 
    - PlanFluor 10x/0.3 DIC
    - PlanFluor 40x/0.75 DIC
    - PlanApo 60x/1.4 oil DIC

    Filters:
    -  GFP, FITC (Ex : 460-500, Dichroic Mirror : 505, BA : 510-560)
    -  Tx Red (Ex : 540-580, DM : 595, BA : 600-660)