Microfluidic immobilization and subcellular imaging of developing Caenorhabditis elegans

Shivers, Jordan and Uppaluri, Sravanti and Brangwynne, Clifford P. (2017) Microfluidic immobilization and subcellular imaging of developing Caenorhabditis elegans. Microfluidics and Nanofluidics, 21 (9). ISSN 1613-4982

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Abstract

Caenorhabditis elegans has been an essential model organism in the fields of developmental biology, neuroscience, and aging. However, these areas have been limited by our ability to visualize and track individual C. elegans worms, especially at the subcellular scale, over the course of their lifetime. Here we present a microfluidic device to culture individual C. elegans in parallel throughout post-embryonic development. The device allows for periodic mechanical immobilization of the worm, enabling 3D imaging at subcellular precision. The immobilization is sufficient to enable fluorescence recovery after photobleaching (FRAP) measurements on organelles and other substructures within the same specific cells throughout larval development, without the use of chemical anesthetics. Using this device, we measure FRAP recovery of two nucleolar proteins in specific intestinal cells within the same worms during larval development. We show that these proteins exhibit different fluorescence recovery as the worm grows, suggesting differential protein interactions during development. We anticipate that this device will help expand the possible uses of C. elegans as a model organism, enabling its use in addressing fundamental questions at the subcellular scale.

Item Type:	Article
Authors:	Shivers, Jordan and Uppaluri, Sravanti and Brangwynne, Clifford P.
Document Language:	Language English
Subjects:	Social sciences > Sociology & anthropology > Communities > Planning and development > Development > Urban development Social sciences > Economics > Production > Economic policy, economic development, economic growth
Divisions:	Azim Premji University - Bengaluru > School of Arts and Sciences
Full Text Status:	Restricted
URI:	http://publications.azimpremjiuniversity.edu.in/id/eprint/7306
Publisher URL:	https://doi.org/10.1007/s10404-017-1988-2

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