Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy.
Submitted by Daniel Alfonso Colón-Ramos on
Title | Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy. |
Publication Type | Journal Article |
Year of Publication | 2013 |
Authors | Wu, Y, Wawrzusin, P, Senseney, J, Fischer, RS, Christensen, R, Santella, A, York, AG, Winter, PW, Waterman, CM, Bao, Z, Colón-Ramos, DA, McAuliffe, M, Shroff, H |
Journal | Nat Biotechnol |
Volume | 31 |
Issue | 11 |
Pagination | 1032-8 |
Date Published | 2013 Nov |
ISSN | 1546-1696 |
Keywords | Animals, Brain, Caenorhabditis elegans, Cell Tracking, Human Umbilical Vein Endothelial Cells, Humans, Imaging, Three-Dimensional, Lighting, Microscopy, 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. |
DOI | 10.1038/nbt.2713 |
Alternate Journal | Nat. Biotechnol. |
PubMed ID | 24108093 |
PubMed Central ID | PMC4105320 |
Grant List | GM097576 / GM / NIGMS NIH HHS / United States HD075602 / HD / NICHD NIH HHS / United States R01 NS076558 / NS / NINDS NIH HHS / United States R01NS076558 / NS / NINDS NIH HHS / United States U01 HD075602 / HD / NICHD NIH HHS / United States U01HD075602 / HD / NICHD NIH HHS / United States |