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Microfluidic immobilization and subcellular imaging of developing Caenorhabditis elegans

Jordan Shivers, Sravanti Uppaluri, Clifford P. Brangwynne
doi: https://doi.org/10.1101/139915
Jordan Shivers
1Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA
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Sravanti Uppaluri
1Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA
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Clifford P. Brangwynne
1Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA
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  • For correspondence: cbrangwy@princeton.edu
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Abstract

C. 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 exhibit different fluorescence recovery as they grow, 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, including the role of phase transitions in driving spatiotemporal intracellular organization within multicellular organisms.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 19, 2017.
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Microfluidic immobilization and subcellular imaging of developing Caenorhabditis elegans
Jordan Shivers, Sravanti Uppaluri, Clifford P. Brangwynne
bioRxiv 139915; doi: https://doi.org/10.1101/139915
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Microfluidic immobilization and subcellular imaging of developing Caenorhabditis elegans
Jordan Shivers, Sravanti Uppaluri, Clifford P. Brangwynne
bioRxiv 139915; doi: https://doi.org/10.1101/139915

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