Hello,
I would like to detect ROS using DHE in tumor sample in situ. I have thought of the following scenario:
- to inject 20 mg/kg DHE to mice ip and after 1 hour to sacrifice the mice, take out the tumor, snap freeze, cut with cryostat and visualize under the microscope.
Do you think it is appropriate for detecting ROS within the tissue?
Any comment, suggestion is appreciated!
Thank you!
Eva
Chapter Measurement of Reactive Oxygen Species (ROS) and Mitochondri...
Article Methods for Detection of Mitochondrial and Cellular Reactive...
https://www.researchgate.net/post/DHE_Dihydroethidium_and_unspecific_ROS_staining
The above readings discuss the method or "pros and cons", especially the last discussion. Good luck.
Hello Eva Korpos
You may refer to a similar protocol given below in which the investigators visualized free radicals by dihydroethidine staining.
In situ detection of ROS production
Dihydroethidine (DHE) (50 mg/ml) in dimethylsulfoxide (Invitrogen) was diluted 1:50 in 0.05 M PBS and maintained at 37 C. At 48hand 24h before killing, mice received an ip injection of 300 μl DHE. At the end of the chronic or acute hyperosmolarity procedure, mice were anesthetized and perfused with 4% paraformaldehyde in 0.1 M phosphate buffer. Frozen brain sections (20 μm) were observed with a Zeiss Axioskop coupled to a Zeiss AxioCam.
For more information you may please refer to the research article attached below.
https://academic.oup.com/endo/article/153/3/1317/2424021
Best.
