Biophysics's area is performing research on different research topics: single molecule biophysics, single cell manipulation and imaging, tissue imaging, neural imaging and biosensing. Manipulation tools, such as optical and magnetic tweezers, which are essential to shed light into some fundamental aspects of biological processes (transcription, protein synthesis, DNA replication, virus infections, cells interaction, exocytosis, muscle contractions, etc.) are used. High sensitivity and high colocalisation experiments are performed to track single molecules with nanometer accuracy by mean of high sensitivity fluorescence detection and active trapping stabilization methods. Optical manipulation methods like laser nanosurgery and micro trapping are employed to investigate particular features in cells and tissues. Nonlinear microscopy techniques including two-photon fluorescence microscopy and second-harmonic generation microscopy are developed and used in ex vivo and in vivo measurements ranging from neuroscience to dermatology. Also, Light-sheet based microscopy techniques are used to study whole brain micron-scale neuroanatomy.

Advanced/Correlative microscopy

The group applies advanced imaging methods to the integrated study of subcellular structure and molecular processes occurring in living cells and organisms. One area of development of the group regards correlative microscopy between optical (confocal, nonlinear, SPIM) and electron microscopy. These methods are applied on isolated cells (for example cardiomyocites) to correlate findings from functional imaging measurements with ultrastructural characterization. At the whole organism level, the group aims at implementing methods for imaging development in small animals (zebrafish) and conduct cell lineage studies.

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Neuronal and Cardiac Imaging

Neuronal and cardiac imaging research team aims to develop innovative imaging methodologies for an increased understanding of biological events in brain and heart. Novel implementations of light-sheet microscopy are applied to resolve neuronal anatomy in whole fixed brains with cellular resolution. Moving to living samples, real-time dynamics of brain rewiring are visualized through two-photon microscopy with the spatial resolution of single synaptic contacts. The plasticity of the injured brain is also dissected through cutting-edge optical methods that specifically ablate single neuronal processes. Finally, random access microscopies in combination with novel fluorescence probes allow optical registrations of action potential and calcium release across population of neurons and cardiomyocytes. The development and the application of these complementary optical methodologies will provide fundamental insights in brain and cardiac disease and will represent a whole new approach for the investigation of the physiology of excitable cells.

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Single Molecule Manipulation and Imaging

Single Molecule research team aims at studying the dynamics of biological macromolecules using laser manipulation and imaging techniques. Our research interest is mainly focused on the influence of force on biological processes, the dynamics and mechanics of molecular motors, and the interaction between proteins and nucleic acids. Manipulation of single molecules is realized using high-resolution optical tweezers, which allow probing sub-nanometer conformational changes with sub-millisecond time resolution. Imaging and localization of single molecules with nanometer accuracy is performed through fluorescent probes and advanced microscopy approaches. The goal is to draw a picture of basic biological processes through the analysis of the dynamics and properties of the individual molecules involved.

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Biomedical Imaging

Biomedical imaging research team aims at developing optical methods for tissue diagnostics using microscopic and spectroscopic techniques. The development and integration of multiple laser scanning imaging techniques provides a high-resolution label-free alternative to standard histopathological methods for tissue diagnostics since it allows integrating morphological and functional information and correlating the observed molecular and cellular changes with disease behavior. Moving to spectroscopy, the implementation of fluorescence, Raman, and reflectance spectroscopy within fiber optic probes allows for a more detailed classification of tissues, providing a more accurate diagnosis in terms of both sensitivity and specificity as well as the potential for endoscopic applications. The methods are applied to a broad range of tissues in collaboration with medical experts.

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Superesolution and single molecule tracking

The last years have witnessed exciting advancements to bypass the limited spatial resolution inherent to standard optical microscopy. New generations of super-resolution microscopes, fluorophores and data analysis methods have allowed reaching resolution of few nanometers. We have set a RESOLFT microscope, which is able to image, based on principles reminiscent of those used for STED microscopy, cellular structures labelled with photo-switchable fluorescent proteins in living cells, with a lateral resolution down to 40 nm. Our team is also performing super-resolution techniques based on single-molecule detection such as PALM and STORM. Single molecule localization methods are used to carry out 2D and 3D tracking in-vitro or in living cells. We are currently challenging problems related to Alzheimer's disease and cancer, and involving membrane rafts, cytoskeleton and membrane proteins.

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