Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy
Yicong Wu,Peter Wawrzusin,Justin Senseney,Robert S. Fischer,Ryan Christensen,Anthony Santella,Andrew York,Peter Winter,Clare M. Waterman,Zhirong Bao,Daniel A. Colón-Ramos,Matthew McAuliffe,Hari Shroff +12 more
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TLDR
A dual-view, plane illumination microscope with improved spatiotemporal resolution is developed by switching illumination and detection between two perpendicular objectives in an alternating duty cycle to study biological systems that require high-speed volumetric visualization and/or low photobleaching.Abstract:
Optimal four-dimensional imaging requires high spatial resolution in all dimensions, high speed and minimal photobleaching and damage. We developed a dual-view, plane illumination microscope with improved spatiotemporal resolution by switching illumination and detection between two perpendicular objectives in an alternating duty cycle. Computationally fusing the resulting volumetric views provides an isotropic resolution of 330 nm. As the sample is stationary and only two views are required, we achieve an imaging speed of 200 images/s (i.e., 0.5 s for a 50-plane volume). Unlike spinning-disk confocal or Bessel beam methods, which illuminate the sample outside the focal plane, we maintain high spatiotemporal resolution over hundreds of volumes with negligible photobleaching. To illustrate the ability of our method to study biological systems that require high-speed volumetric visualization and/or low photobleaching, we describe microtubule tracking in live cells, nuclear imaging over 14 h during nematode embryogenesis and imaging of neural wiring during Caenorhabditis elegans brain development over 5 h.read more
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Tissue clearing and its applications in neuroscience
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TL;DR: How tissue-clearing methods could provide an unbiased, system-level view of mammalian bodies and human specimens is discussed and future opportunities for the use of these methods in human neuroscience are discussed.
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TL;DR: Super-resolution optical microscopy is an important tool in a number of scientific disciplines including chemical, (bio)physical and biomedical research, particularly with respect to the study of living cells and organisms.
References
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Journal ArticleDOI
The structure of the nervous system of the nematode Caenorhabditis elegans
TL;DR: The structure and connectivity of the nervous system of the nematode Caenorhabditis elegans has been deduced from reconstructions of electron micrographs of serial sections as discussed by the authors.
Journal ArticleDOI
The embryonic cell lineage of the nematode Caenorhabditis elegans.
TL;DR: It is concluded that the cell lineage itself, complex as it is, plays an important role in determining cell fate and is demonstrated to demonstrate substantial cell autonomy in at least some sections of embryogenesis.
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Bayesian-Based Iterative Method of Image Restoration
TL;DR: An iterative method of restoring degraded images was developed by treating images, point spread functions, and degraded images as probability-frequency functions and by applying Bayes’s theorem.
Journal ArticleDOI
An iterative technique for the rectification of observed distributions
TL;DR: In this article, the authors consider the problem of determining the distribution of proper motions in a line-of-sight line of sight (LoSOS) image from a given number count.
Journal ArticleDOI
Microtubule polymerization dynamics
TL;DR: This review describes progress toward understanding the mechanism of dynamic instability of pure tubulin and discusses the function and regulation of microtubule dynamic instability in living cells.
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