[DIYbio] Re: How does MitoTracker Bind?

Hi, I wonder if you have figured out why MitoTracker specifically binds to mitochondria?

I've been thinking and searching and don't have much clue.

Thanks.

On Saturday, October 24, 2009 2:30:08 PM UTC-5, phillyj wrote:

On Fri, Oct 23, 2009 at 1:52 PM, Nathan McCorkle <nmz...@gmail.com> wrote:
>
> Take a look at these, I think once it gets into the mito it gets
> metabolized and then it stays there... Not sure, I can look more at
> the last link in this list in a few hours, but gotta go to a thesis
> defense on 3d protein visualisation now
>
> http://bbc.mcw.edu/Computation/models/text/Mitochondrial%20Inner%20Membrane%20Electrophysiology%20Assessed%20by%20Rhodamine-123%20Transport%20and%20Fluorescence.pdf
>
I'm reading this one right now. Not a quick reader though.

I sent an email to my professor about what I learned so far and he replied:

" The odd thing is that mitotracker Green-FM (which is the one we
used) seems to have somewhat different properties from the others,
since it appears to stain mitochondria on dead cells --such as cheek
smears, that are dried down before staining. It may be that some
oxidative activity survives the treatment, but it seems unlikely.
Note also that in the cheek cells, it survives despite the lack of
fixation.   It also seems to have a significant background membrane
staining, sugesting that there is a lipid component to the reaction.

In past years, we have been unable to get clear mitotracker staining
on the chicken cells.  This year, we modified the procedure, following
the ideas suggested by the Invitrogen information, and we added the
mitotracker solution to the living cultures prior to aldehyde
fixation.  As you saw, there was very little stain in the fixed cells,
and there were major problems with cell attachment and survival. There
were also many odd circular patches with punctate fluorescence all
over the slides.  This stain clearly had nothing to do with the cells.
 My suspicion is that it represents some form of micelle that
developed in the culture medium and then bound to the glass.
Interestingly, when we looked at the chicken cells before fixation,
but after mitotracker exposure, we did see convincing mitochondrial
stain, but all of the cells were floating.  Bummer.

As you can see, there seems to be more to the Mitotracker Green-FM
than is in the Invitrogen description.  In fact, if you compare the
literature in the "manual" with the "handbook", you will find some
inconsistencies between the two sources."

According to him, Mitotracker green FM is not like the other ones. I
wonder what's going on.

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