Time-lapse imaging of Drosophila testis for monitoring actin dynamics and sperm release

Tushna Kapoor; Pankaj Dubey; Krishanu Ray

doi:10.1016/j.xpro.2021.101020

Time-lapse imaging of Drosophila testis for monitoring actin dynamics and sperm release

STAR Protoc. 2021 Dec 15;3(1):101020. doi: 10.1016/j.xpro.2021.101020. eCollection 2022 Mar 18.

Authors

Tushna Kapoor¹, Pankaj Dubey¹, Krishanu Ray¹

Affiliation

¹ Department of Biological Sciences, Tata Institute of Fundamental Research, Mumbai, Maharashtra 400005, India.

Abstract

Here we describe a simple step-by-step protocol for collecting high-resolution, time-lapse images of intact Drosophila testis ex vivo for a limited period using a confocal microscope, with minimum photo-toxic damage, to monitor spermatid individualization, coiling, and release. The F-actin dynamics during spermatid morphogenesis can be further investigated through laser ablations, Fluorescence-Recovery-After-Photobleaching, and drug treatments, using this protocol. For complete details on the use and execution of this protocol, please refer to Dubey et al. (2016), Dubey et al. (2019), and Kapoor et al. (2021).

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actins*
Animals
Drosophila*
Male
Spermatozoa
Testis
Time-Lapse Imaging

Substances

Actins