Grant: Research Tools and Instruments - Category 1 (<$150,000)

High-throughput protein immunolocalization analysis through virtual fluorescence imaging.The objective of this grant proposal is to acquire an immunocytolocalization numeric fluorescent module to complement and expand the range of analyses provided by the Faculty of Medicine and Health Sciences histology and electronic microscopy core (hereafter named Histology Core). The research services provided by the Histology Core is already available to the entire Faculty of medicine research teams, and the number of users is steadily increasing. The addition of a fluorescent module to our Hamamatsu Nanozoomer 2.0-RS slide scanner, obtained through CFI (No 20939), will enable multi-channel fluorescence scanning and digital numerization of whole slides following protein fluorescent immunolocalization either from histological tissues or fixed cells, including yeasts. Contrary to conventional microscopy, this state-of-the-art apparatus reduces the area of light excitation thus reducing the lost of fluorescent signals through photobleaching while allowing the detection and digital numerization of faint fluorescent signals. The Nanozoomer series is equipped with the Z-stack feature that allows focusing at different depths in 3D samples such as clumps of cells and thick tissues. The virtual image resulting from the Nanozoomer scan provides a picture of the whole slide and not a selected area, thus reducing possible bias that could be introduced by the observer. Hence, the digitalized virtual image allowed researchers to zoom on a specific region without losing image resolution or having to go back for further microscopic observations. In fact, the requested system will greatly expand our capabilities by providing high content non-biased imaging system that accurately digitizes object slides at the highest possible fluorescence resolution, over a complete range of standard magnifications, therefore offering panoramic mapping, screening and multispectral histocytometric image analysis of whole tissue sections.