Measurement of Lysosomal Size and Lysosomal Marker Intensities in Adult Caenorhabditis elegans

Julie M Huynh; Hope Dang; Hanna Fares

doi:10.21769/BioProtoc.2724

Measurement of Lysosomal Size and Lysosomal Marker Intensities in Adult Caenorhabditis elegans

Bio Protoc. 2018 Feb 5;8(3):e2724. doi: 10.21769/BioProtoc.2724.

Authors

Julie M Huynh¹, Hope Dang², Hanna Fares²

Affiliations

¹ College of Medicine-Tucson, University of Arizona, Tucson, USA.
² Department of Molecular and Cellular Biology, University of Arizona, Tucson, USA.

Abstract

Assays have been developed to study trafficking in various tissues of Caenorhabditis elegans. Adult C. elegans intestinal cells are large and have extensive endocytic networks, thus making them a good system for deciphering the endocytic pathway using live imaging techniques. However, the presence of auto-fluorescent gut granules in adult intestine can interfere with the signals of endocytic compartment reporters, like GFP. Here we demonstrate a protocol adapted from the original method developed by the Grant laboratory to identify signals from reporters in adult intestinal cells. The goal of this protocol is to identify endocytic compartments tagged with fluorescent markers without any confounding effects of background autofluorescent gut granules in adult intestinal cells of Caenorhabditis elegans.

Keywords: C. elegans; Confocal microscopy; Endocytosis; Gut granule; Intestine; Lysosome.