Ovarial dissections and staining

Sonja Fritzsche; Vera S. Hunnekuhl

Improve Research Reproducibility A Bio-protocol resource

Home
Protocols

Concise Method

Ovarial dissections and staining

SF Sonja Fritzsche

VH Vera S. Hunnekuhl

This method is extracted from research article: EvoDevo, Jun 2021

Cell-specific expression and individual function of prohormone convertase PC1/3 in Tribolium larval growth highlights major evolutionary changes between beetle and fly neuroendocrine systems

DOI: 10.1186/s13227-021-00179-w

Ask a question

Favorite

Ovaries were dissected out of the abdomina from 14–15-day old females. A subset of ovaries classified into the respective phenotypes were further processed for fluorescent staining. The ovarioles were individually freed from their muscular lining with ultra-fine dissection needles. Ovaries were then fixed in 4% formaldehyde/ PBT for 30 min and stained overnight in Phalloidin-A488 (Sigma) at a concentration of 1:40 (v/v in PBT). Specimens were then washed 4 × 10 min in PBT and stained with DAPI (v/v 1/1000 of a 1 µg/µl stock solution in PBT) for 20 min. After 2 more PBT washes ovaries were mounted and imaged in the same way described for the stained nervous systems above.

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

Post a Question

0 Q&A

Share your protocol with your peers.

Submit a Preprint Protocol