Inverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans.

Imagen de Daniel Alfonso Colón-Ramos
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TítuloInverted selective plane illumination microscopy (iSPIM) enables coupled cell identity lineaging and neurodevelopmental imaging in Caenorhabditis elegans.
Publication TypeJournal Article
Year of Publication2011
AutoresWu, Y, Ghitani, A, Christensen, R, Santella, A, Du, Z, Rondeau, G, Bao, Z, Colón-Ramos, DA, Shroff, H
JournalProc Natl Acad Sci U S A
Date Published2011 Oct 25
Palabras claveAnimals, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Cell Lineage, Homeodomain Proteins, Imaging, Three-Dimensional, Microscopy, Nervous System, Time Factors

The Caenorhabditis elegans embryo is a powerful model for studying neural development, but conventional imaging methods are either too slow or 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 30× 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.

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID22006307
PubMed Central IDPMC3203761
Grant List5 T32GM07499-34 / GM / NIGMS NIH HHS / United States
F32 GM091874 / GM / NIGMS NIH HHS / United States
R00 HG004643 / HG / NHGRI NIH HHS / United States
R00 NS057931 / NS / NINDS NIH HHS / United States