Selective stains for Mitochondria (fixed tissue)..

[Bernoull]

---

Members don't see this ad.

Can someone guide me towards resources (protocols/publications/books)
on selectively staining mitochondria in fixed tissue (human brain & rabbit, rat cardiac tissue)??

I'm looking for relatively simple but effective methods. I've found excellent literature on immunohistochemistal (IHC) methods, but I can't use IHC right now so I'm thinking of chemical stains. I know of Cain's method but it doesn't seem to be selective enough...

Any thoughts would really be appreciated!!!

Thanks

Mods: I posted this is the Allo forum to gain from people's background in histology/pathology etc...

 

[BrainPathology]

Can someone guide me towards resources (protocols/publications/books)
on selectively staining mitochondria in fixed tissue (human brain & rabbit, rat cardiac tissue)??

I'm looking for relatively simple but effective methods. I've found excellent literature on immunohistochemistal (IHC) methods, but I can't use IHC right now so I'm thinking of chemical stains. I know of Cain's method but it doesn't seem to be selective enough...

Any thoughts would really be appreciated!!!

Thanks

Mods: I posted this is the Allo forum to gain from people's background in histology/pathology etc...

Found this:

http://www.histosearch.com/histonet/Oct99A/Re.MITOCHONDRIALSTAIN.html

My lab techs haven't ever done any of the techniques listed on that list but if you have the resources you might try it. I'm thinking you may have come across this already in your own search though. It was relatively quick to find with a google search.

My own texts on mitochondrial stains assume cryostat tissue so none of them would be any help to you. If you ever DO have access to frozen tissue I'd definitely be interested in how an NADH, SDH or COX enzyme histochemistry looks on tissue other than muscle.

 

[Bernoull]

Found this:

http://www.histosearch.com/histonet/Oct99A/Re.MITOCHONDRIALSTAIN.html

My lab techs haven't ever done any of the techniques listed on that list but if you have the resources you might try it. I'm thinking you may have come across this already in your own search though. It was relatively quick to find with a google search.

My own texts on mitochondrial stains assume cryostat tissue so none of them would be any help to you. If you ever DO have access to frozen tissue I'd definitely be interested in how an NADH, SDH or COX enzyme histochemistry looks on tissue other than muscle.

Many thanks for helping out!!

I actually never saw that specific thread but I've seen another thread on Cain's Method from that site. I'll dig some more about the methods they suggested and hopefully find a protocol somewhere..

For now, I'll start out with Cain's method. Do you have any suggestions about how I can find literature that evaluates the pros/cons & effectiveness of the various mitochondrial staining methods (cain's, altmann's)? Would this likely be in a text or research publication? This would save me a lot of time/money or trial/error work..

Would you recommend any journals and texts on histological staining? I found a copy of the Hist. staining by Armed Force Institute of Pathology but it's quite dated from 1960!!

Finally is histochemistry applicable to paraffin sections or do the preservation techniques render paraffin section useless for histochemistry?

My apologies for all the questions, but I really appreciate whatever insights you have!!

Thanks again

I may have access to cryostat sections, and I'll let you know what I find with respect to MT staining in cardiac/brain tissues.

 

[BrainPathology]

Many thanks for helping out!!

I actually never saw that specific thread but I've seen another thread on Cain's Method from that site. I'll dig some more about the methods they suggested and hopefully find a protocol somewhere..

For now, I'll start out with Cain's method. Do you have any suggestions about how I can find literature that evaluates the pros/cons & effectiveness of the various mitochondrial staining methods (cain's, altmann's)? Would this likely be in a text or research publication? This would save me a lot of time/money or trial/error work..

Would you recommend any journals and texts on histological staining? I found a copy of the Hist. staining by Armed Force Institute of Pathology but it's quite dated from 1960!!

Finally is histochemistry applicable to paraffin sections or do the preservation techniques render paraffin section useless for histochemistry?

My apologies for all the questions, but I really appreciate whatever insights you have!!

Thanks again

I may have access to cryostat sections, and I'll let you know what I find with respect to MT staining in cardiac/brain tissues.

My understanding (and since I'm not actually a histotech I'm going by what I was just told) is that formalin and enzyme histochemistry are mutually exclusive. My elementary understanding of formalin fixation is that it irreversibly binds amino groups with any nearby nitrogen atoms in whatever molecule is available, whether that be DNA or other proteins etc. (any organic chemist is probably snickering at how basic that is though)

That AFIP book is still used by a lot of labs including the one where I trained. These stains don't really get updated and a 2010 book would likely just repeat everything done in the 60s except the way reagents are disposed of and would likely not encourage use of certain chemicals that impractical in a lot of places becaus of environmental laws (we had to stop using B5 because of mercury for example). If that book has a protocol I would try it. I don't know what your background is but if it's not in histology I would definitely go to your histo lab and ask for help and insight there. Now that you actually have an idea of the names of protocols you want to use you have a starting point. Some reagents are notoriously finicky and that may help you decide with them which one might be more successful.

No idea about journals specific to this though. Your head histotech would be a much better resource for this as well.

IF you do get access to frozen tissue and your lab does muscle biopsy enzyme histochemistry the NADH, SDH, and COX stains will be routine to them. You will effectively eliminate the variable of having to learn how to make a new protocol (for your lab) work. OR you'll have great control staining to evaluate how well the protocols on the paraffin tissue is going.

Best of luck. I'd really be curious how you solve this in the end.