Three-dimensional modeling and reconstruction for multimodality phase microscopy

Phase microscopy modalities are extensively used to image living transparent biological samples because of their ability to obtain high contrast images without the use of enhancing agents. However when the imaged object is optically thick, that is, when the thickness of the object is larger than the depth of field of the imaging system, the development of relevant image modeling and reconstruction techniques is necessary to extract useful information from the images.;The contribution of this work is toward the development of both a theoretical model and reconstruction techniques for phase images of optically thick objects. In particular, we have developed and tested a generative forward model based on a product of convolutions (POC) approach for these phase images. The POC model is based on the use of the point spread function (PSF) of the optical system and can be used to simulate phase images of any transparent object geometry. We have also developed an initial reconstruction method, specifically a multi-modal boundary-constrained inversion that combines two distinct phase imaging modalities, differential interference contrast (DIC) and optical quadrature microscopy (OQM ) modalities. We use DIC to extract 3D information about object morphology and OQM to obtain a quantitative phase image from the entire object. The reconstruction algorithm combines the information from the two modalities to obtain the spatial variation of the indices of refraction of the imaged objects.;Our methods could be used to improve the analysis of a number of classes of biological and biomedical samples. We use as a prototype application mouse embryo development studies, where the goal is to extract detailed information about the embryo at different development stages. We present results showing validation of the work done to date using simulated and measured multimodality phase images of both synthetic and mouse embryo samples.