Light Sheet Microscopy: From Molecules to Organism Imaging
1Research Center for Applied Science, Academia Sinica, Taipei, Taiwan
* presenting author:Bi-Chang Chen,
We crafted a massively parallel array of coherently interfering beams composing a two-dimensional optical lattices [1]. To apply such light sheets to in vivo imaging, the lattice pattern will be conjugated to the rear pupil plane of excitation objective and an orthogonal detection objective will be suspended from above with their ends dipped in a shallow media-filled and temperature controlled bath. As the specimen is moved through the light sheet, the fluorescence thereby generated is recorded as series of 2D images on a camera. These are then assembled into a 3D image, and the process is repeated to build a 4D data set of cellular dynamics. We operated the microscope in one of two different modes: a suppuration SIM mode or a high-speed dithered mode. The technology collects high-resolution images rapidly and minimizes damage to cells, meaning it can image the three-dimensional activity of molecules, cells, and embryos in fine detail over longer periods than was previously possible. We could image three-dimensional (3D) dynamics for hundreds of volumes, often at sub-second intervals, at the diffraction limit and beyond.

Keywords: light-sheet microscopy , fluorescence microscopy, 3D live imaging