Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans Journal Article
| Authors: | Wu, Y.; Ghitani, A.; Christensen, R.; Santella, A.; Du, Z.; Rondeau, G.; Bao, Z.; Colón-Ramos, D.; Shroff, H. |
| Article Title: | Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans |
| Abstract: | The Caenorhabditis elegans embryo is a powerful model for studying neural development, but conventional imaging methods are either too slowor phototoxic to take full advantage of this system. To solve these problems, we developed an inverted selective plane illumination microscopy (iSPIM) module for noninvasive high-speed volumetric imaging of living samples. iSPIM is designed as a straightforward add-on to an inverted microscope, permitting conventional mounting of specimens and facilitating SPIM use by development and neurobiology laboratories. iSPIM offers a volumetric imaging rate 30x faster than currently used technologies, such as spinning-disk confocal microscopy, at comparable signal-to-noise ratio. This increased imaging speed allows us to continuously monitor the development of C, elegans embryos, scanning volumes every 2 s for the 14-h period of embryogenesis with no detectable phototoxicity. Collecting ∼25,000 volumes over the entirety of embryogenesis enabled in toto visualization of positions and identities of cell nuclei. By merging two-color iSPIM with automated lineaging techniques we realized two goals: (i) identification of neurons expressing the transcription factor CEH-10/Chx10 and (ii) visualization of their neurodevelopmental dynamics. We found that canal-associated neurons use somal translocation and amoeboid movement as they migrate to their final position in the embryo. We also visualized axon guidance and growth cone dynamics as neurons circumnavigate the nerve ring and reach their targets in the embryo. The high-speed volumetric imaging rate of iSPIM effectively eliminates motion blur from embryo movement inside the egg case, allowing characterization of dynamic neurodevelopmental events that were previously inaccessible. |
| Keywords: | controlled study; unclassified drug; microscopy; nonhuman; neuroimaging; animals; confocal microscopy; embryo; molecular dynamics; signal noise ratio; transcription factor; embryo development; homeodomain proteins; molecular imaging; automation; time factors; cell lineage; imaging, three-dimensional; cell migration; cell cycle arrest; caenorhabditis elegans; nervous system development; nervous system; volumetry; caenorhabditis elegans protein; caenorhabditis elegans proteins; illumination; bacterial cell; nerve fiber growth; axon growth; fast 4d imaging; neuron migration; transcription factor ceh 10; biological monitoring; inverted selective plane illumination microscopy; neurobiology |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 108 |
| Issue: | 43 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2011-10-25 |
| Start Page: | 17708 |
| End Page: | 17713 |
| Language: | English |
| DOI: | 10.1073/pnas.1108494108 |
| PROVIDER: | scopus |
| PMCID: | PMC3203761 |
| PUBMED: | 22006307 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 9 December 2011" - "CODEN: PNASA" - "Source: Scopus" |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work