Mitochondria theoretical home experiment idea

MAOAr297r

MAOAr297r

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Hey everyone. This might be really dumb and I may be really far off but I'd like to hear you're thoughts. I really want to perform this in my amateur/student lab soon.

 
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PinkPanda

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I must say, I have a bit trouble judging if this is a good idea and could work or not, I don't know much about working with a microscope and the technical stuff sorrounding it.

Maybe, if you have the energy, you could write a very short text on your experimental setup? I'm not a native speaker and I didn't get all the technical terms.

Otherwhise, if you manage to try your experiment, please update how it goes. I'm still trying to understand how it works, but I'd be interested in the results! :D
 
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Hutan

Hutan

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PinkPanda said:
I must say, I have a bit trouble judging if this is a good idea
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Same. Is the test a known thing? I imagine it might be done in lots of classrooms. So the only unknown is how fast the change happens in both you and a control?

But I applaud the effort when it's clearly costing you a lot to think it all through and make the video.

Good luck and I look forward to hearing how you go.

My temperature is always a bit low and I've heard others with ME/CFS find the same thing. I'm not sure how universal it is though. I wonder how much that amount of temperature depression affects the respiration processes or if it is the low rate of energy use that affects the temperature (or some circular combination). I suppose there would be some data on the rate of respiration at different temperatures.

I guess you would have to be aware of the temperature of the two samples when you look at them under the microscope - maybe the light might heat up the slide more in the second sample?

And I guess, there is the possibility that our cheek cells might be fine while our brain cells or immune system cells are the ones going slow?

My brain has packed up for the night. Hope there was something coherent there.

But anyway, the main thing was, well done. You clearly are not taking this illness lying down (except for the literal lying down bit).
 
Hip

Hip

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That sounds like a very interesting idea and experiment. It would be great if this approach using the janus green B dye allowed one to simply and directly measure the hypo-metabolism in ME/CFS patient's cells.

I found this video of cheek cells under the microscope being stained with the janus green B dye.

And this video (at 4:46) shows a microscopic close up of a cell's mitochondria made visible by the janus green B stain. A screenshot of this video is shown below:

Mitochondria Stained Blue Using Janus Green B Dye
Mitochondria made visible by staining with janus green B.png

Janus green B can be bought: here for $47, here for $16, here for $6, and here for $30.
 
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Hip

Hip

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This paper mentions that janus green B is reduced by the lactic dehydrogenase and the glucose dehydrogenase enzyme systems, but does not mention the effect of pyruvate dehydrogenase on this stain.
 
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Wishful

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Hutan is correct about cheek cell mitochondria not necessarily being useful for researching ME/CFS. The brain's mitochondria are genetically different from those in the rest of the body, so we could have problems with one but not the other.

My 'not feeling worse than usual' oral temperature is 36.65C. Yes, that's probably overly precise, but several mercury thermometers all agreed on it being between 36.6 and 36.7 most of the time. Top 'feeling extremely lousy' temperature was 37.6C. Typical 'feeling lousy' temperature is 36.8-37.0C for a fairly short time.
 
Hip

Hip

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Wishful said:
Hutan is correct about cheek cell mitochondria not necessarily being useful for researching ME/CFS.
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Fluge and Mella showed that in cultured human myoblast cells not even belonging to the ME/CFS patient, when they exposed those myoblast cells in vitro to the serum of ME/CFS patients, the cells developed energy metabolism abnormalities.

So that suggests that "there is something in the serum", and that in ME/CFS patients, cells in contact with the serum (which is nearly all cells in the body, including epithelial cells from the cheek) might develop energy hypo-metabolism.

MBM used a completely different type of cell to demonstrate energy hypo-metabolism in ME/CFS: neutrophils from the blood of ME/CFS patients.
 
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MAOAr297r

MAOAr297r

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Hey everyone! Thank you for all the encouragement, the help and the kind support. I successfully conducted the beginning of this today. I should have a result by friday or so. I'm going to get some help from a retired clinical microbiologist this week.

I wish I could get well enough to go back to school so I could get the education I need to really know what I'm doing. I'm double majoring in molecular bio and neuroscience. It's so frustrating sometimes. I crave the proper education that would allow to me actually study this but I'm just too sick to function, it's so frustrating. I want to get educated and join the fight on the biological front so bad :( I'm sooooo curious.
 
tiredowl

tiredowl

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Hip said:
Fluge and Mella showed that in cultured human myoblast cells not even belonging to the ME/CFS patient, when they exposed those myoblast cells in vitro to the serum of ME/CFS patients, the cells developed energy metabolism abnormalities.

So that suggests that "there is something in the serum", and that in ME/CFS patients, cells in contact with the serum (which is nearly all cells in the body, including epithelial cells from the cheek) might develop energy hypo-metabolism.

MBM used a completely different type of cell to demonstrate energy hypo-metabolism in ME/CFS: neutrophils from the blood of ME/CFS patients.
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Something in the serum that's not belonging to ME/CFS patients? Could someone elaborate on what this mean?
Does that mean we are contagious?
 
Hip

Hip

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tiredowl said:
Something in the serum that's not belonging to ME/CFS patients? Could someone elaborate on what this mean?
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It pretty much explains it in my post: if you take healthy muscle cells, and expose them to the blood serum of ME/CFS patients, those muscle cells will rapidly develop energy metabolism abnormalities.

So that suggests that in the blood of ME/CFS patients, there is some factor which interferes with the energy metabolism. That factor is unknown, but could be for example some sort of signaling molecule that actually modulates the cellular energy metabolism. Or it could be some autoantibody that disrupts energy metabolism. By energy metabolism we mean the functioning of things like mitochondria and glycolysis.

It's not contagious, as this factor is not thought to be a pathogen.
 
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