Light Sheet-based Fluorescence Microscopy (LSFM) reduces Phototoxic Effects by several Orders of Magnitude and provides new Means for the Modern Life Sciences
Prof. Dr. Ernst Stelzer (Goethe-Universität Frankfurt)
In conventional and confocal fluorescence microscopy, recording stacks of images along the optical z-axis illuminates the entire specimen once for each plane. Hence, cells are illuminated 10-20 and fish embryos 100-300 times more often than they are observed. This can be avoided by using light sheets, which are fed into the specimen from the side and overlap with the focal plane of a wide-field fluorescence microscope. In contrast to an epi-fluorescence arrangement, an azimuthal fluorescence arrangement uses two independently operated lenses for illumination and detection. Optical sectioning and no photo-toxic damage or photo-bleaching outside a small volume close to the focal plane are intrinsic properties.