title: Long-Term High-Resolution Imaging of Developing C. elegans Larvae with Microfluidics.
abstract: Long-term studies of Caenorhabditis elegans larval development traditionally require tedious manual observations because larvae must move to develop, and existing immobilization techniques either perturb development or are unsuited for young larvae. Here, we present a simple microfluidic device to simultaneously follow development of ten C. elegans larvae at high spatiotemporal resolution from hatching to adulthood (∼3 days). Animals grown in microchambers are periodically immobilized by compression to allow high-quality imaging of even weak fluorescence signals. Using the device, we obtain cell-cycle statistics for C. elegans vulval development, a paradigm for organogenesis. We combine Nomarski and multichannel fluorescence microscopy to study processes such as cell-fate specification, cell death, and transdifferentiation throughout post-embryonic development. Finally, we generate time-lapse movies of complex neural arborization through automated image registration. Our technique opens the door to quantitative analysis of time-dependent phenomena governing cellular behavior during C. elegans larval development.
authors.affiliation: Center for Physics and Biology, The Rockefeller University, New York, NY 10065, USA; Laboratory of Developmental Genetics, The Rockefeller University, New York, NY 10065, USA. Electronic address: wkeil@rockefeller.edu.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Keil
authors.affiliation: Laboratory of Developmental Genetics, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Lena M
authors.initials: LM
authors.lastname: Kutscher
authors.affiliation: Laboratory of Developmental Genetics, The Rockefeller University, New York, NY 10065, USA. Electronic address: shaham@rockefeller.edu.
authors.forename: Shai
authors.initials: S
authors.lastname: Shaham
authors.affiliation: Center for Physics and Biology, The Rockefeller University, New York, NY 10065, USA. Electronic address: siggiae@rockefeller.edu.
authors.forename: Eric D
authors.initials: ED
authors.lastname: Siggia
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D002453:Cell Cycle; D016923:Cell Death; D002455:Cell Division; D058948:Cell Tracking; D054338:Cell Transdifferentiation; D005260:Female; D018507:Gene Expression Regulation, Developmental; D021621:Imaging, Three-Dimensional; D007814:Larva / Q000378:metabolism; D008297:Male; D044085:Microfluidics / Q000379:methods*; D016501:Neurites / Q000378:metabolism; D013997:Time Factors; D059008:Time-Lapse Imaging; D014844:Vulva / Q000166:cytology / Q000254:growth & development
keywords: Caenorhabditis elegans; PDA neuron; PVD neuron; dendritic arborization; larval development; linker cell death; long-term imaging; microfluidics; transdifferentiation; vulval development