Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Phalloidin-Hoechst staining of cells

SS Sarbani Saha
SC Shreeta Chakraborty
AB Agnihotri Bhattacharya
AB Arati Biswas
RA Rupasri Ain
request Request a Protocol
ask Ask a question
Favorite

To visualize the formation of syncytium, BeWo cells were grown on coverslip in a 35 mm dish in the absence and presence of 8-Bromo-cAMP and fixed with freshly prepared ice-cold 4% para-formaldehyde for 15 minutes at room temperature followed by three PBS washes (5 minutes each). To stain the cytoskeleton, fixed cells were incubated with phalloidin (Cell Signaling Technology) at 1:200 dilution for 15 minutes. Then the cells were again washed with PBS for three times and incubated with Hoechst at a final concentration of 2 mg/ml for another 15 minutes to stain the nuclei. Then the cells were rinsed with PBS for several times and finally, the coverslip was mounted on a glass slide with the help of fluoroshield (Sigma, USA) and examined under Leica epifluorescence microscope.

Copyright and License information: The Author(s) ©2017
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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, 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.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A