TY - JOUR
T1 - Fast objective coupled planar illumination microscopy
AU - Greer, Cody J.
AU - Holy, Timothy E.
N1 - Funding Information:
We thank Zhongsheng Guo, Dae Woo Kim, and Ben Acland for their work in developing imaging software and the Washington University Medical School Machine Shop for their help in designing and machining custom optomechanical parts (John Kreittler, John Witte, and Kevin Poenicke). We also thank the users and staff of the Washington University Center for Cellular Imaging for consistent feedback that enabled us to debug and improve the microscope. Finally, we thank Xiaoyan Fu, Donghoon Lee, and Terra Barnes for comments and suggestions regarding the paper. This work was supported by NIH grants 1R24NS086741-01, 5R01NS068409-08, and 1T32NS073547-01.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Among optical imaging techniques light sheet fluorescence microscopy is one of the most attractive for capturing high-speed biological dynamics unfolding in three dimensions. The technique is potentially millions of times faster than point-scanning techniques such as two-photon microscopy. However light sheet microscopes are limited by volume scanning rate and/or camera speed. We present speed-optimized Objective Coupled Planar Illumination (OCPI) microscopy, a fast light sheet technique that avoids compromising image quality or photon efficiency. Our fast scan system supports 40 Hz imaging of 700 μm-thick volumes if camera speed is sufficient. We also address the camera speed limitation by introducing Distributed Planar Imaging (DPI), a scaleable technique that parallelizes image acquisition across cameras. Finally, we demonstrate fast calcium imaging of the larval zebrafish brain and find a heartbeat-induced artifact, removable when the imaging rate exceeds 15 Hz. These advances extend the reach of fluorescence microscopy for monitoring fast processes in large volumes.
AB - Among optical imaging techniques light sheet fluorescence microscopy is one of the most attractive for capturing high-speed biological dynamics unfolding in three dimensions. The technique is potentially millions of times faster than point-scanning techniques such as two-photon microscopy. However light sheet microscopes are limited by volume scanning rate and/or camera speed. We present speed-optimized Objective Coupled Planar Illumination (OCPI) microscopy, a fast light sheet technique that avoids compromising image quality or photon efficiency. Our fast scan system supports 40 Hz imaging of 700 μm-thick volumes if camera speed is sufficient. We also address the camera speed limitation by introducing Distributed Planar Imaging (DPI), a scaleable technique that parallelizes image acquisition across cameras. Finally, we demonstrate fast calcium imaging of the larval zebrafish brain and find a heartbeat-induced artifact, removable when the imaging rate exceeds 15 Hz. These advances extend the reach of fluorescence microscopy for monitoring fast processes in large volumes.
UR - http://www.scopus.com/inward/record.url?scp=85072916549&partnerID=8YFLogxK
U2 - 10.1038/s41467-019-12340-0
DO - 10.1038/s41467-019-12340-0
M3 - Article
C2 - 31578369
AN - SCOPUS:85072916549
SN - 2041-1723
VL - 10
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 4483
ER -