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Proceedings Paper

Adaptive optics microscopy enhances image quality in deep layers of CLARITY processed brains of YFP-H mice
Author(s): Marc R. Reinig; Samuel W. Novack; Xiaodong Tao; Florian Ermini; Laurent A. Bentolila; Dustin G. Roberts; Allan MacKenzie-Graham; S. E. Godshalk; M. A. Raven; Joel Kubby
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Paper Abstract

Optical sectioning of biological tissues has become the method of choice for three-dimensional histological analyses. This is particularly important in the brain were neurons can extend processes over large distances and often whole brain tracing of neuronal processes is desirable. To allow deeper optical penetration, which in fixed tissue is limited by scattering and refractive index mismatching, tissue-clearing procedures such as CLARITY have been developed. CLARITY processed brains have a nearly uniform refractive index and three-dimensional reconstructions at cellular resolution have been published. However, when imaging in deep layers at submicron resolution some limitations caused by residual refractive index mismatching become apparent, as the resulting wavefront aberrations distort the microscopic image. The wavefront can be corrected with adaptive optics. Here, we investigate the wavefront aberrations at different depths in CLARITY processed mouse brains and demonstrate the potential of adaptive optics to enable higher resolution and a better signal-to-noise ratio. Our adaptive optics system achieves high-speed measurement and correction of the wavefront with an open-loop control using a wave front sensor and a deformable mirror. Using adaptive optics enhanced microscopy, we demonstrate improved image quality wavefront, point spread function, and signal to noise in the cortex of YFP-H mice.

Paper Details

Date Published: 9 March 2016
PDF: 12 pages
Proc. SPIE 9690, Clinical and Translational Neurophotonics; Neural Imaging and Sensing; and Optogenetics and Optical Manipulation, 969008 (9 March 2016); doi: 10.1117/12.2213283
Show Author Affiliations
Marc R. Reinig, Univ. of California, Santa Cruz (United States)
Samuel W. Novack, Univ. of California, Santa Cruz (United States)
Xiaodong Tao, Univ. of California, Santa Cruz (United States)
Florian Ermini, Univ. of California, Santa Cruz (United States)
Laurent A. Bentolila, Univ. of California, Los Angeles (United States)
Dustin G. Roberts, Univ. of California, Los Angeles (United States)
Allan MacKenzie-Graham, Univ. of California, Los Angeles (United States)
S. E. Godshalk, Univ. of California, Santa Barbara (United States)
M. A. Raven, Univ. of California, Santa Barbara (United States)
Joel Kubby, Univ. of California, Santa Cruz (United States)


Published in SPIE Proceedings Vol. 9690:
Clinical and Translational Neurophotonics; Neural Imaging and Sensing; and Optogenetics and Optical Manipulation
Steen J. Madsen; E. Duco Jansen; Samarendra K. Mohanty; Nitish V. Thakor; Qingming Luo; Victor X. D. Yang, Editor(s)

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