Etude de l'effet de la variation d'indice de réfraction par modélisation de la fonction d'étalement du point (PSF) en microscopie de flurescence

Abstract

In fluorescence microscopy, the thickness of the specimen is high relative to the depth of field of a high numerical aperture (Na) objective of the imaging system. An aberration is introduced when there is a variation in the refractive index in the specimen. Aberrations decrease the resolution and contrast of the image. Deconvolution is a method used to restore the image by computation and numerical simulation, and for an efficient deconvolution, an accurate knowledge of the point spread function (PSF) of the imaging system is essential. This PSF can be measured experimentally or modeled theoretically. Experimentally, the PSF can be obtained by capturing 3D images of a point source of size smaller than the resolution of the microscope objective. But it is often tedious to prepare an accurate experimental setup. Therefore, a theoretical model that takes into account the optical parameters and imaging properties to model the PSF is a good alternative. In addition, this PSF model must take into account the aberrations, in order to conform to the experimental conditions.