Optical effects of the cranium in trans-cranial in-vivo two photon laser scanning microscopy in mice

P. Johannes Helm; Ole P. Ottersen M.D.; Gabriele Nase

doi:10.1117/12.700241

12 February 2007 Optical effects of the cranium in trans-cranial in vivo two photon laser scanning microscopy in mice

P. Johannes Helm, Ole P. Ottersen M.D., Gabriele Nase

Proceedings Volume 6442, Multiphoton Microscopy in the Biomedical Sciences VII; 644221 (2007) https://doi.org/10.1117/12.700241
Event: SPIE BiOS, 2007, San Jose, California, United States

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.

Citation Download Citation

P. Johannes Helm, Ole P. Ottersen M.D., and Gabriele Nase "Optical effects of the cranium in trans-cranial in vivo two photon laser scanning microscopy in mice", Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 644221 (12 February 2007); https://doi.org/10.1117/12.700241

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