In order to study ROS and mitochondria morphology I require to study these things
Thank you guys in advance.
Hello Yogesh,
in the attachment you will find an excellent paper on changes of mitochondrial morphology during ageing. It contains some interesting EM studies.
-> Morpho-dynamic changes of mitochondria during ageing of human endothelial cells by Jendrach et al., 2005 published in Mech. Ageing Dev.
Best regards,
Christian
Hello Yogesh
In my opinion you have several techniques instead of EM to study mitochondrial morphology. At EM level you can see changes in cristae and density but to quantify any parameter you would need a lot of images to avoid variability. If you want to address changes in mitochondrial morphology or ROS, I think is better to use confocal or fluorescence microscopy and comercial probes (mitotrackers, mitoSOX....) or antibodies
Good luck
David
Hello Yogesh,
Please find attached our 2010 AJP paper: Morphological and functional abnormalities in mitochondria associated with synaptic degeneration in prion disease.
I believe it provides a good guidance for investigations of mitochondrial morphology using transmission electron microscopy. Using this approach we examined a wide range of mitochondrial structural parameters, including inner membrane ultrastructure. I think you may be able to benefit from reading this study.
Feel free to contact me if you need any further advice!
Best,
Zuzana
Hi there,
Similarly, we investigated mitochondrial size, shape and density in an optic nerve model of neurotrauma, where we know oxidative stress is a significant factor. heres the link:
http://www.biomedcentral.com/1471-2202/14/98 (open access)
and a paper with classic oxidative stress morphology in TBI which I found helpful:
http://www.nature.com/jcbfm/journal/v26/n11/full/9600297a.html
however, they did the EM on isolated mitochondria
Good luck!
In our experience, EM is a excelent method for studying mitochondrial morphology changes, but fixation is a critical step. Mitochondria is the first organelle suffering artifacts during fixation. In animal research, perfusion fixation respecting volume/intravascular pression is the best.
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