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Long-Term High-Resolution Imaging of Developing C. elegans Larvae with Microfluidics

Cornell Affiliated Author(s)

Author

W. Keil
L.M. Kutscher
S. Shaham
E.D. Siggia

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. © 2017 Elsevier Inc.

Date Published

Journal

Developmental Cell

Volume

40

Issue

2

Number of Pages

202-214,

URL

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85009383585&doi=10.1016%2fj.devcel.2016.11.022&partnerID=40&md5=d7d8c0773253054274afe79cddc58d26

DOI

10.1016/j.devcel.2016.11.022

Research Area

Funding Source

ECCS-1542081
HD078703
NS064273
NS081490
PHY 1502151
LT000250/2013-C
P40 OD010440
R01HD078703
GM066699

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