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Proceedings Paper

Optical effects of the cranium in trans-cranial in vivo two photon laser scanning microscopy in mice
Author(s): P. Johannes Helm; Ole P. Ottersen; Gabriele Nase
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Paper Abstract

The combination of multi photon laser scanning microscopy with transgenic techniques has set the stage for in vivo studies of long term dynamics of the central nervous system in mice. Brain structures located within 100?m to 200?m below the brain surface can be observed minimum-invasively during the post-adolescent life of the animal. However, even when selecting the most appropriate microscope optics available for the purpose, trans-cranial observation is compromised by the aberrations induced by the cranium and the tissue interposed between the cranium and the actual focus. It still is an un-resolved task to calculate these aberrational effects or to, at least, estimate quantitatively the distortions they induce onto the recorded images. Here, we report about measurements of the reflection, the absorption, and the effects on the objective point spread function of the mouse cranium as a function of the thickness of the cranium, the locus of trans-cranial observation and the wavelength. There is experimental evidence for pronounced Second Harmonic Generation (SHG) effects.

Paper Details

Date Published: 12 February 2007
PDF: 9 pages
Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 644221 (12 February 2007); doi: 10.1117/12.700241
Show Author Affiliations
P. Johannes Helm, Univ. of Oslo (Norway)
Ole P. Ottersen, Ctr. for Molecular Biology and Neuroscience, Univ. of Oslo (Norway)
Gabriele Nase, Ctr. for Molecular Biology and Neuroscience, Univ. of Oslo (Norway)


Published in SPIE Proceedings Vol. 6442:
Multiphoton Microscopy in the Biomedical Sciences VII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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