Measuring the acidification of the phagosomal lumen in live C. elegans embryos

Omar Peña-Ramos; Zheng Zhou

doi:10.1016/j.xpro.2023.102332

Measuring the acidification of the phagosomal lumen in live C. elegans embryos

STAR Protoc. 2023 Jun 3;4(2):102332. doi: 10.1016/j.xpro.2023.102332. Online ahead of print.

Authors

Omar Peña-Ramos¹, Zheng Zhou²

Affiliations

¹ Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA.
² Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, USA. Electronic address: zhengz@bcm.edu.

Abstract

In metazoans, the acidification of the phagosomal lumen is essential for the efficient degradation of cargoes. Here, we present a protocol for measuring the rate of acidification inside phagosomal lumen containing apoptotic cells in living C. elegans embryos. We describe steps for generating a worm population, selecting embryos, and mounting embryos on agar pads. We then detail live imaging of embryos and data analysis. This protocol is applicable to any organism in which real-time fluorescence imaging can be performed. For complete details on the use and execution of this protocol, please refer to Pena-Ramos et al. (2022).¹.

Keywords: Cell Biology; Developmental Biology; Microscopy; Model Organisms.

Grants and funding

R01 GM067848/GM/NIGMS NIH HHS/United States