Isotropic light-sheet microscopy and automated cell lineage analyses to catalogue caenorhabditis elegans embryogenesis with subcellular resolution Review
| Authors: | Duncan, L. H.; Moyle, M. W.; Shao, L.; Sengupta, T.; Ikegami, R.; Kumar, A.; Guo, M.; Christensen, R.; Santella, A.; Bao, Z.; Shroff, H.; Mohler, W.; Colón-Ramos, D. A. |
| Review Title: | Isotropic light-sheet microscopy and automated cell lineage analyses to catalogue caenorhabditis elegans embryogenesis with subcellular resolution |
| Abstract: | Caenorhabditis elegans (C. elegans) stands out as the only organism in which the challenge of understanding the cellular origins of an entire nervous system can be observed, with single cell resolution, in vivo. Here, we present an integrated protocol for the examination of neurodevelopment in C. elegans embryos. Our protocol combines imaging, lineaging and neuroanatomical tracing of single cells in developing embryos. We achieve long-term, four-dimensional (4D) imaging of living C. elegans embryos with nearly isotropic spatial resolution through the use of Dual-view Inverted Selective Plane Illumination Microscopy (diSPIM). Nuclei and neuronal structures in the nematode embryos are imaged and isotropically fused to yield images with resolution of ~330 nm in all three dimensions. These minute-by-minute high-resolution 4D data sets are then analyzed to correlate definitive cell-lineage identities with gene expression and morphological dynamics at single-cell and subcellular levels of detail. Our protocol is structured to enable modular implementation of each of the described steps and enhance studies on embryogenesis, gene expression, or neurodevelopment. © 2019 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. |
| Keywords: | microscopy; nonhuman; gene expression; embryo; developmental biology; embryo development; publication; cell lineage; caenorhabditis elegans; visualization; nervous system development; lineage; neurodevelopment; illumination; elegans; embryos; identity; c; deconvolution; article; cytoshow; dispim; issue 148; spimfusion; starrynite |
| Journal Title: | Journal of Visualized Experiments |
| Volume: | 2019 |
| Issue: | 148 |
| ISSN: | 1940-087X |
| Publisher: | MYJoVE Corporation |
| Date Published: | 2019-06-01 |
| Start Page: | e59533 |
| Language: | English |
| DOI: | 10.3791/59533 |
| PUBMED: | 31233035 |
| PROVIDER: | scopus |
| PMCID: | PMC7255390 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 2 August 2019 -- Source: Scopus |
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