Time-lapse and cleared imaging of mouse embryonic lung explants to study three-dimensional cell morphology and topology dynamics

Harold Fernando Gómez; Nikolaos Doumpas; Dagmar Iber

doi:10.1016/j.xpro.2023.102187

Time-lapse and cleared imaging of mouse embryonic lung explants to study three-dimensional cell morphology and topology dynamics

STAR Protoc. 2023 Mar 22;4(2):102187. doi: 10.1016/j.xpro.2023.102187. Online ahead of print.

Authors

Harold Fernando Gómez¹, Nikolaos Doumpas², Dagmar Iber³

Affiliations

¹ Department of Biosystems Science and Engineering (D-BSSE), ETH Zürich, Basel, Switzerland; Swiss Institute of Bioinformatics (SIB), Basel, Switzerland. Electronic address: harold.gomez001@gmail.com.
² Department of Biosystems Science and Engineering (D-BSSE), ETH Zürich, Basel, Switzerland; Swiss Institute of Bioinformatics (SIB), Basel, Switzerland.
³ Department of Biosystems Science and Engineering (D-BSSE), ETH Zürich, Basel, Switzerland; Swiss Institute of Bioinformatics (SIB), Basel, Switzerland. Electronic address: iberd@ethz.ch.

Abstract

Here, we present a protocol for collecting high-spatiotemporal-resolution datasets of undisturbed mouse embryonic epithelial rudiments using light-sheet fluorescence microscopy. We describe steps for rudiment dissection, clearing, and embedding for cleared and live imaging. We then detail procedures for light-sheet imaging followed by image processing and morphometric analysis. We provide protocol variations for imaging both growing and optically cleared lung explants to encourage the quantitative exploration of three-dimensional cell shapes, cell organization, and complex cell-cell dynamics. For complete details on the use and execution of this protocol, please refer to Gómez et al. (2021).¹.

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