Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

TMRE (mitochondrial membrane potential) staining and microscopy

XL Xiaotong Li
JK Jason Karpac
request Request a Protocol
ask Ask a question
Favorite

For midgut TMRE (tetramethylrhodamine, ethyl ester) staining, dissected in 1×PBS and fixed for 10 min at room temperature in gut fixation solution (100 mM Glutamic Acid, 25 mM KCl, 20 mM MgSO4, 4 mM Na2HPO4, 1 mM MgCl2; pH adjusted to 7.5 with 10 N NaOH, 4% Formaldehyde). Then incubated in 200 nm TMRE staining solution (Abcam, #113852) for 20 min at room temperature. After staining, samples were then rinsed once in wash solution (25 nm TMRE) for 30 s. Samples were quickly mounted in the wash solution onto a slide, kept overnight at 4 °C and imaged using identical setting on the confocal microscope. Confocal images were collected using a Nikon Eclipse Ti confocal system (utilizing a single focal plane) and processed using the Nikon software and Adobe Photoshop.

Copyright and License information: The Author(s) ©2023
Open Access This 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/.

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 Post a Question
0 Q&A