Non-linear micrascopy of biophysical processes in living cells

点击翻译按钮获取中文摘要

Optical microscopy is among the most powerful tools that enable to investigate non-invasively living cells and biological tissue. The proposed new non-linear optical microscope features the shaping unit that is able to manipulate spectral and spatial profiles of the laser beam, as well as adjust the temporal phasing and polarization of different spectral components of the femtosecond pulse. The custom shaped femtosecond pulses will be used for coherent control excitation of fluorophores in the biological samples. The optimized laser pulses will enhance the efficiency of fluorophore excitation and reduce photobleaching. The femtosecond pulse chirp modulation will enable to image the pH distribution inside the cells. The coherent control microscope will be built on the basis of already existing state-of-the-art multimodal non-linear laser scanning microscope capable of simultaneously recording third and second harmonic generation, and multiphoton excitation fluorescence images. The unique instrument will be used to investigate bioenergetics of mitochondria during contraction of myocytes. The microscope will also be applied to elucidate regulation mechanisms of light harvesting in single isolated and in situ chloroplasts. The new microscope will serve as a platform for creation of new non-linear imaging modalities for non-invasive biological investigations.