abstract: During the past two decades, biological imaging has undergone a revolution in the development of new microscopy techniques that allow visualization of tissues, cells, proteins and macromolecular structures at all levels of resolution. Thanks to recent advances in optics, digital sensors and labeling probes, one can now visualize sub-cellular components and organelles at the scale of a few dozens nanometers to several hundreds of nanometers. As a result, fluorescent microscopy and multimodal imaging has become the workhorse of modern biology. All these technological advances in microscopy, created new challenges for researchers in quantitative image processing and analysis. Therefore, dedicated efforts are necessary to develop and integrate cutting-edge approaches in image processing and optical technologies to push the limits of the instrumentation and to analyze the large amount of data being produced.
In this talk, we present image processing methods, mathematical models, and algorithms to build an integrated imaging approach that bridges the resolution gaps between the molecule and the whole cell, in space and time. The presented statical methods are dedicated to the analysis of proteins in motion inside the cell, with a special focus on Rab protein trafficking observed in time-lapse confocal microscopy or total internal reflection fluorescence microscopy. Nevertheless, the proposed image processing methods and algorithms are flexible in most cases, with a minimal number of control parameters to be tuned. They can be applied to a large range of problems in cell imaging and can be integrated in generic image-based workflows, including for high content screening applications.