Faculty Opinions recommendation of SPED light sheet microscopy: fast mapping of biological system structure and function.

The goal of understanding living nervous systems has driven interest in high-speed and large fieldof-view volumetric imaging at cellular resolution. Light-sheet microscopy approaches have emerged for cellular-resolution functional brain imaging in small organisms such as larval zebrafish, but remain fundamentally limited in speed. Here we have developed SPED light sheet microscopy, which combines large volumetric field-of-view via an extended depth of field with the optical sectioning of light sheet microscopy, thereby eliminating the need to physically scan detection objectives for volumetric imaging. SPED enables scanning of thousands of volumes-persecond, limited only by camera acquisition rate, through the harnessing of optical mechanisms that normally result in unwanted spherical aberrations. We demonstrate capabilities of SPED microscopy by performing fast sub-cellular resolution imaging of CLARITY mouse brains and cellular-resolution volumetric Ca2+ imaging of entire zebrafish nervous systems. Together, SPED light sheet methods enable high-speed cellular-resolution volumetric mapping of biological system structure and function. Correspondence: deissero@stanford.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AUTHOR CONTRIBUTIONS R.T. developed the SPED light sheet microscopy, and with K.D. designed the experiments. R.T. and M.L-B. performed the imaging experiments. R.T. analyzed the data. I.K. and R.T. performed the PSF simulations with input from S.Y. S.S. contributed scripts for analysis. L.G., M.B., S.Y. and A.A. led the light field microscopy development. A.A. and V.B. performed the LFM imaging, and contributed to empirical PSF quantification. R.T. and K.D. wrote the paper with editorial input from all authors. K.D. supervised all aspects of the work. Published as: Cell. 2015 December 17; 163(7): 1796–1806. H H M I A uhor M anscript H H M I A uhor M anscript H H M I A uhor M anscript