In vivo imaging of development with combined Optical Tomography and Light Sheet Microscopy

Optical Tomography is a three dimensional imaging technique which, similarly to x-ray computed tomography, is based on the acquisition of a sequence of optical transmission (or fluorescence) images of the sample from several orientations. The acquired images, or projections, are combined to reconstruct the 3D volume of the sample, typically using a backprojection algorithm. The seminar describes recent developments for in-vivo Optical Tomography, towards high resolution imaging of the anatomy of translucent biological samples. Novel contrast mechanisms, based on blood cell motion and muscular tissue birefringence are discussed, together with their applications in developmental biology. The combination of Optical Tomography with Selective Plane Illumination Microscopy (SPIM) is then described. Optical tomography was integrated with SPIM in order to image zebrafish embryos over several hours of development, to detect, segment, track and automatically register single organs during the course of long term time lapse acquisitions.