B16: Nonlinear optical microscopy
Julien Douady, Davy Cottet, Jean-Claude Vial
Laboratoire de Spectrométrie Physique
http://www-lsp.ujf-grenoble.fr/-Microscopies-intravitalesKeywords:
nonlinear optics, two-photon excitation fluorescence, second-harmonic generation,
multi-photon microscopy.
Abstract:
The Laboratoire de Spectrométrie Physique offers a nonlinear optical
microscopy facility, including simultaneous two-photon and second-harmonic
imaging. The applications are devoted to deep microscopy in turbid media (biological
tissues) and furthermore in-vivo imaging in the small animal.
Because of the wavelengths used range in the near-IR, the diffusion
phenomena is reduced ; in addition, due to the nonlinear effect, the fluorescence and
second-harmonic signals are generated only in the focal volume and thus offers an
intrisic 3D-resolution.
Vascularisation in the living mouse brain
Structure of an arterial wall
The experimental work proposed will address the following points :
- characteristics of femto-seconds lasers ;
- scanning microscopy setup ;
- comparison between linear and non-linear response (2-photon fluorescence) ;
- second-harmonic signal : effect of the anisotropic emission ;
- model of a diffusive media and advantages of nonlinear methods ;
- imaging of some biological tissues.
Note that the relative importance of each part will depend on the background of the
students.
A revue on this topic, including other lectures suggestions:
http://www-lsp.ujf-grenoble.fr/IMG/html/NEN.html