Have I been working with tissues and cell culture, have not found a lot of information of Mitosox in tissue, I know can be used, so he wanted to know if someone has experience?
Hello Yenddy,
please have a look at the publication in the attachment. The authors used MitoSOX staining in cardiac tissue samples.
I hope this information is useful for you.
Regards,
Christian
Article Succinate dehydrogenase inhibition with malonate during repe...
If the question is can MitoSOX be used in histological slices, I would say no because mROS has a very short half-life. However, tissue samples, such as the ones in the paper suggestion by Christian would probably work fine under the right stimulation. I would be curious about tissue penetration, however, if the tissue were not dissociated before measurement.
Dear Yenddy,
In the way we used in the article sent by Christian, MitoSOX Red worked relatively well. We first fixed our samples in paraformaldehid, and then we incubated them with MitoSOX Red 5 microM, for 15 min at room temperature in the dark. After washing, we also stained nuclei with Hoechst. However, in the discussion of our article we mentioned that mitoSOX staining after fixation is in fact measuring total ROS instead of mitochondrial ROS. In the discussion of our article we said: "In order to measure mitochondrial ROS production in cardiac tissue, MitoSOX staining should be performed before tissue fixation. However, in cardiac tissue, this approach presents several technical limitations that preclude its use, particularly the exposure of the samples to ischaemia while they are incubated with the probe. For this reason, we performed MitoSOX staining after fixation. This method provides us information on total rather than mitochondrial ROS production."
Thank you to everyone for their valuable help, try to make it in tissues, including in OCT without fixing them, Thanks a lot . Yen
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