Expansion-assisted selective plane illumination microscopy for nanoscale imaging of centimeter-scale tissues
Adam K. Glaser, Jayaram Chandrashekar, Sonya Vasquez, Cameron Arshadi, Naveen Ouellette, Xiaoyun Jiang, Judith Baka, Gábor Kovács, Micah Woodard, Sharmishtaa Seshamani, Kevin Cao, Nathan Clack, Andrew Recknagel, Anna Grim, Pooja Balaram, Emily Turschak, Alan Liddell, John R. Rohde, Ayana Hellevik, Kevin Takasaki, Lindsey A. Barner, Molly Logsdon, Chris Chronopoulos, Saskia de Vries, Jonathan T. Ting, Steve I. Perlmutter, Brian Kalmbach, Nikolai Dembrow, R. Clay Reid, David Feng, Karel Svoboda
Abstract
Abstract Recent advances in tissue processing, labeling, and fluorescence microscopy are providing unprecedented views of the structure of cells and tissues at sub-diffraction resolutions and near single molecule sensitivity, driving discoveries in diverse fields of biology, including neuroscience. Biological tissue is organized over scales of nanometers to centimeters. Harnessing molecular imaging across three-dimensional samples on this scale requires new types of microscopes with larger fields of view and working distance, as well as higher imaging throughput. We present a new expansion-assisted selective plane illumination microscope (ExA-SPIM) with diffraction-limited and aberration-free performance over a large field of view (85 mm2) and working distance (35 mm). Combined with new tissue clearing and expansion methods, the microscope allows nanoscale imaging of centimeter-scale samples, including entire mouse brains, with diffraction-limited resolutions and high contrast without sectioning. We illustrate ExA-SPIM by reconstructing individual neurons across the mouse brain, imaging cortico-spinal neurons in the macaque motor cortex, and tracing axons in human white matter.