Mounting Drosophila pupae for laser ablation and live imaging of the dorsal thorax

James T O'Connor; Erica K Shannon; M Shane Hutson; Andrea Page-McCaw

doi:10.1016/j.xpro.2022.101396

Mounting Drosophila pupae for laser ablation and live imaging of the dorsal thorax

STAR Protoc. 2022 May 14;3(2):101396. doi: 10.1016/j.xpro.2022.101396. eCollection 2022 Jun 17.

Authors

James T O'Connor^{1

2}, Erica K Shannon^{1

2}, M Shane Hutson^{3

4

5}, Andrea Page-McCaw^{1

2

6}

Affiliations

¹ Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN, USA.
² Program in Developmental Biology, Vanderbilt University, Nashville, TN, USA.
³ Department of Physics and Astronomy, Vanderbilt University, Nashville, TN, USA.
⁴ Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA.
⁵ Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, USA.
⁶ Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, TN, USA.

Abstract

This protocol describes the preparation of Drosophilamelanogaster pupae for laser ablation and live imaging of the notum (dorsal thorax). Because the pupa is stationary, it can be continuously live imaged for multiple days if desired, making it ideal for studying wound signaling and repair, from before laser ablation through wound closure. In this protocol, we demonstrate the processes of staging, partially dissecting, mounting, wounding, and live imaging the pupal notum, with the wounding occurring during the live imaging process. For complete details on the use and execution of this protocol, please refer to O'Connor et al. (2021b).

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

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
Drosophila melanogaster
Drosophila*
Laser Therapy*
Pupa
Thorax / diagnostic imaging

Abstract

Publication types

MeSH terms

Grants and funding