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Counting Mitochondria

Mitochondria

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#1 Ozhan Sean

Ozhan Sean

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Posted 22 January 2016 - 12:52 PM

Hello everyone! I am new to this forum! 

Okay, straight to it. I have been trying to find good ways to count and measure mitochondria in my epithelial cells, extracted through mouth swap, and track the average over time. I could use mDNA ratio etc to keep a track of increase or decrease of mitochondria in cells over time. I have the capacity to do PCR and electrophoresis and all that, but I cant isolate cells, cant make sure I am getting the same number of cells, and there are a few other variables I cant control. Even if I did I would still not learn about the number and size of mitochondria. I really dont want to spend the money to get mitotracker dyes, they aren't cheap, and pics others have taken online don't seem like they would help in counting, they just highlight an area rich with mitochondria. I thought I would just use methelyn blue dye and look for them under the scope, then I would use ImageJ to count them. But then what I found was that the cells had only 20 to 30 mitochondria after dye, which is strange because I expected a lot more. Secondly the ImageJ software was not helpful at all. I would count 30, but the software would say 600.

Here is one of the the pics I took. I am curious: 

1) Am I really looking at mitochondria in these pictures? Shouldn't there be more? They are shaped just like mitochondria, but they seem TOO large, and too few. 

2) To get a good average I will need to count the mitochindria in a bunch of cells, that's a lot of work for a party of one, to do every day! Specially if I want to get an average measure of the size of mitochondria. That is time I don't have, anyone know a better software or a method? 

3) Has anyone here done this? Any ideas?

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