A system for automated high-content screening and laser manipulation of single cells.

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Fluorescence microscopy is an essential technology that is widely used to study a myriad of cellular processes. Numerous fluorescent probes have been designed to visualize and analyze a large range of molecules, from ions to nucleic acids and proteins. These probes are commonly used in cells to report the presence and localization of molecules in both fixed and live cells, as well as tissue and whole organisms. Fluorescence microscopes, thus, are fundamental tools to understand the intricate intracellular interactions between different molecules and proteins, by simply analysing light emitted by these engineered probes.******The applicants currently use this technology for a wide range of applications on a daily basis, where individual images are used to characterize the behaviour of tens or hundreds of cells. Nevertheless, several critical studies require the number of cells to be much higher (e.g. testing the effects of several drugs in parallel in many different conditions). For this, we are requesting a motorized system that can image millions of cells for extended periods of time in large cell culture chambers. This feature requires the microscope to be able to automatically refocus the sample before images are obtained. However, most systems do this by processing images, by analysing different features of the photos with algorithms, but this approach does not provide the accuracy we require. The system we are applying for uses an internal laser that is reflected on the cell substrate and provides precise information about the position of the sample to maintain focus. The automation of positioning and refocusing allows interrogation of fine details of cell behaviour with high resolution in a large number of cells and conditions. ******Finally, we need to use external lasers to manipulate cells individually. For example, we may search for cells that exhibit a predefined phenotype and illuminate only those cells. The configuration of the system that we request allows integration/addition of external laser systems that we already have in place. These external ports that enable adaptation and expansion for novel technology developments are unique features for which we have a pressing need and cannot be obtained as a turn-key solution. This infrastructure will also allow training of highly qualified personnel in the development of innovative engineering solutions to solve biological questions.********