Visualization of large scale objects using confocal microscope leica TSC LSI

• Autorzy: Platek R. , Korczynski J. , Skup M.
• Języki publikacji: EN

Abstrakty

EN

Conventional confocal microscopy is dedicated mainly to proceed with thin tissue sections and cells seeded on a cover slip. The Leica TCS LSI macro confocal is the first super zoom microscope that combines all benefits of traditional confocal microscopy with large scale imaging of anesthetized, alive objects as well as unfixed/fixed objects post mortem. In our experiments we have tested Leica TCS LSI for scanning of (1) murine brains and spinal cords in vivo, and dissected in toto immediately post mortem, and (2) fixed rat spinal cords. We used transgenic mice expressing green fluorescent protein (GFP) under PLP promoter, to visualize oligodendrocytes, and rats with spinal cords transduced with AAV vector coding for enhanced GFP under mCMV promoter, to visualize neurons and glia. Super zoom confocal microscopy let us observe general distribution of GFP-expressing cells in whole organs as well as to focus on single cells and fibers. We were able to discriminate well between main morphological features of these cells. In murine brains we could visualize myelinated axons and oligodendrocytes, whereas in rat spinal cords the extent of eGFP expressing cells and their fibers traversing along entire spinal cord could be traced. The labeled objects could be visualized from the regions lying within a range of 150 - 350 μm from the surface of the brain/spinal cord. The details on the procedures applied, benefits and limitations of the method will be presented.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2011
• Tom: 71
• Numer: S
• Opis fizyczny: p.13

Twórcy

autor

Platek R.

• Laboratory for Reinnervation Processes, Department of Neurophysiology, Nencki Institute of Experimental Biology PAS, Warsaw, Poland

autor

Korczynski J.

• Laboratory for Confocal Microscopy, Nencki Institute of Experimental Biology PAS, Warsaw, Poland

autor

Skup M.

• Laboratory for Reinnervation Processes, Department of Neurophysiology, Nencki Institute of Experimental Biology PAS, Warsaw, Poland

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