Light Sheet Fluorescence Microscopy (LSFM)

Light sheet microscopy utilizes a planar illumination technique to provide a unique way of volumetric imaging for thin (<10um) samples like cultured cells as well as for thick specimens like tissues prepped with various clearing methods. The imaging technique combines a detecting lens installed orthogonally to one or two illuminating lenses with better penetration capability into deep samples (low NA). By illuminating only the section around a focal plane, light sheet technique offers a significant reduction in out-of-focus blur, photobleaching, and phototoxicity compared to conventional widefield and confocal microscopy. These features enable to track live samples over an extended period of time in a more physiological condition, which benefits many fields including neurobiology and developmental biology. One of the difficulties in choosing a LSFM comes from the variety of geometries that are available. You should pay attention to the orientation of the illumination and detection light paths in relationship to the sample and limitations on sample mounting. 


Reference for Clearing Methods

iBiology - Light Sheet Sectioning by Dr. Ernst Stelze