Dear Andreia,

just recently I read about the relatively new method called PpIX-TSLT (Protoporohyrin IX - Triplet State Lifetime Technique). It's mainly a medical approach to meassure mitochondrial oxygen tension through the skin. But perhaps it's something you could use.

Here is a paper regarding this method:

https://ccforum.biomedcentral.com/articles/10.1186/s13054-015-1056-9

Best regards,

Chris

Thanks for your suggestion Chris!

I don't know if it would be aplicable to my studies but I will take a look

Well. We (laboratory of structure and functions of mitochondria) use oxygen electrode and confocal microscopy mostly.

So if you have good confocal (or very good fluorescent) microscope, which can resolve mitochondria - you can work with different fluoresent probes. For potential (tmre), ros (dcf), different mitotracers and so on. If you can resolve individual mitochondria you can do quite a lot of things - measure quantity, do 3d segmentation (if you have z-stacks) and have their volume, check their movement and shape (fragmentation process). And you can check dynamics of this processes and reaction to some changes be it chemical, or physical (like oxygen depletion). You can try to fixate and use immunohistochemistry.

Plus you can isolate individual mitochondria and use flow cytometry on functioning mito or check something via western blotting or similar methods.

Much depends on your microscope and object.

Hope it helps, I can try to think more if you specify task a little.

Thank you Vasily! Very nice suggestions. Indeed I was thinking on using microscopy for assessing the structure and fragmentation. I hope our microscope has enough resolution. My question was really more related to the measurement of respiration and such parameters but as far as I could find we really need to use the oxygen electrode.

I use mitochondrial enzymatic activities to determine mitochondrial function. Depending on what your objective is it is a fairly cost effective way of getting a lot of data.

Tyler Batchelder Andreia N Carvalho Vasily A Popkov Chris Samek I very much appreciate the topic and comments. As a clinician I get the general idea of what you are all saying, but have a question about context. I'm not clear if these methods are for whole-body mitochondrial function, or if they are tissue-specific. We are developing a biophysics-based technology to enhance tissue function and reduce inflammation. We are looking for a way to assess mitochondrial function in in-vivo or in-vitro tissues. For instance, looking at muscle or chondrocyte or tendon metabolomics in a person, and/or looking at metabolomic/mitochondrial function in in-vivo tissue samples. Is that possible with any of the techniques you're discussing? What direction should I go in to explore this?

Hi!

We have been using Extracellular Flux Analyzer to measure oxygen consumptiom rate, maybe this is useful for your experiments.