A nanoscopy platform for live cell and tissue imaging based on adaptive illumination and spatial light modulation

The accurate subcellular localization of proteins, organelles (mitochondria, endoplasmatic reticulum) and cellular extensions (immune cell filopodia, sensory nerve endings) is critically important for correct functioning of many different cells. Mislocalization can cause discomfort (eg. chronic pain), slowly developing (eg. neurodegeneration, cancer) or acute life threatening (eg. cardiac) diseases. We can study the spatial organization in living cells with fluorescence imaging, however the resolution of classic microscopy is limited due to diffraction, which blurs the image and prevents separation of two points in close vicinity. Substantial effort was put in solving this problem and superresolution techniques were developed successfully, for which the pioneers received in 2014 the Nobel prize for Chemistry (E. Betzig, S. Hell, W.E. Moerner). The microcopy platform applied for in this proposal uses the principle of STED (stimulated emission depletion), one of the superresolution approaches that is fast and can be used in living cells. The scientists in charge of this project all have questions that need this type of resolution to advance their research concerning: pain, neurodegeneration, cancer, cardiac and gut diseases and bacterial toxin production. We chose to work with new electro-optical elements (spatial light modulation) and adaptive illumination, as developed in the lab of S. Hell, to assure that nanoscale imaging is possible in living cells and complex tissues.