Dual-view plane illumination microscopy for rapid and spatially isotropic imaging Journal Article
| Authors: | Kumar, A.; Wu, Y.; Christensen, R.; Chandris, P.; Gandler, W.; McCreedy, E.; Bokinsky, A.; Colón-Ramos, D. A.; Bao, Z.; McAuliffe, M.; Rondeau, G.; Shroff, H. |
| Article Title: | Dual-view plane illumination microscopy for rapid and spatially isotropic imaging |
| Abstract: | We describe the construction and use of a compact dual-view inverted selective plane illumination microscope (diSPIM) for time-lapse volumetric (4D) imaging of living samples at subcellular resolution. Our protocol enables a biologist with some prior microscopy experience to assemble a diSPIM from commercially available parts, to align optics and test system performance, to prepare samples, and to control hardware and data processing with our software. Unlike existing light sheet microscopy protocols, our method does not require the sample to be embedded in agarose; instead, samples are prepared conventionally on glass coverslips. Tissue culture cells and Caenorhabditis elegans embryos are used as examples in this protocol; successful implementation of the protocol results in isotropic resolution and acquisition speeds up to several volumes per s on these samples. Assembling and verifying diSPIM performance takes ∼6 d, sample preparation and data acquisition take up to 5 d and postprocessing takes 3-8 h, depending on the size of the data. |
| Keywords: | adult; young adult; microscopy; nonhuman; animal cell; image analysis; embryo; validation study; imaging system; image quality; imaging; caenorhabditis elegans; collimator; equipment design; image processing; image display; tissue culture cell; optical resolution; computer analysis; agarose; process development; camera; data processing; image retrieval; article; imaging software; dual view plane illumination microscopy; isotropic imaging; parasite examination |
| Journal Title: | Nature Protocols |
| Volume: | 9 |
| Issue: | 11 |
| ISSN: | 1754-2189 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2014-11-01 |
| Start Page: | 2555 |
| End Page: | 2573 |
| Language: | English |
| DOI: | 10.1038/nprot.2014.172 |
| PROVIDER: | scopus |
| PUBMED: | 25299154 |
| PMCID: | PMC4386612 |
| DOI/URL: | |
| Notes: | Export Date: 1 December 2014 -- Source: Scopus |
