Eosin Y with Methylene Blue?
Eosin Y with Methylene Blue?
Hello! I picked up some Methylene Blue from an online seller. It also came with a bottle of Eosin Y, and the pamphlet that came with it describes it as a counterstain to Methylene Blue. Is that true? I can't find a procedure online for using it as a counterstain to Methylene Blue. I was under the impression that the two stains were largely used for different specimens, with Methylene Blue more effective on animal nuclei, and Eosin Y useful for plant cytoplasm. Also, when I've used these stains before, Methylene Blue was effective immediately, while Eosin Y needed some soaking time, so I'm not sure what the procedure would be.
Re: Eosin Y with Methylene Blue?
Hi, well I haven't used this combination myself before although I've used and have both of them. My use is entirely for Botanical sectioned and mounted, sometimes 'live', tissue.
What these 2 are though is, on paper a reasonable pairing. The MB is as you say a nuclear stain that also may be used as a 'general stain' as it will stain all tissue-types, or virtually all (again, in the context of plant tissue). EY is again as said, primarily a cytoplasmic stain - the two together in a tissue will give a nice figure with good contrast, of blue/black nuclei against the light-pink cell contents.....
EY will take ages to stain plant tissue however, while MB must be used both very dilute and relatively quickly - as it is a 'bit of an animal' when it comes to staining - a very, very enthusiastic stain!
Consider them to be a classic stain/counterstain combination and go from there, perhaps by reference to another such combination such as Safranin(nuclear) with alcian-blue(cytoplasmic), or indeed Safranin(nuclear) with the excellent fast-green as the cellulosic and cytoplasmic counterstain.....
The basic method is, nuclear-stain first, remove excess with alcohol or water or a mixture of both, to isolate (differentiate) the nuclear-stain to the nuclei ideally. Then apply the counterstain to 'stain the rest' as it were, but apply this as an 'additive' stain - i.e. stain, examine, stain some more, examine etc until you have the desired figure....
A huge subject - you may try each stain on it's own first to see how it performs and is used, before serious attempts at the bi-stains.
My advice would be to forget the EY - I rarely find it satisfactory.
Use the MB as a nice general-stain and buy qalcian-blue, safranin and fast-green, in powder form, from e-bay. They're all very cheap to buy and work very well (and permanently) together..... Remember however that the above is in the context of Botanical staining.....
Search internet for safranin+fast-green staining protocol, and safranin+alcian-blue protocol - in fact if you search this forum I very likely have several posts re this with images that will help you a lot....
Good luck.
What these 2 are though is, on paper a reasonable pairing. The MB is as you say a nuclear stain that also may be used as a 'general stain' as it will stain all tissue-types, or virtually all (again, in the context of plant tissue). EY is again as said, primarily a cytoplasmic stain - the two together in a tissue will give a nice figure with good contrast, of blue/black nuclei against the light-pink cell contents.....
EY will take ages to stain plant tissue however, while MB must be used both very dilute and relatively quickly - as it is a 'bit of an animal' when it comes to staining - a very, very enthusiastic stain!
Consider them to be a classic stain/counterstain combination and go from there, perhaps by reference to another such combination such as Safranin(nuclear) with alcian-blue(cytoplasmic), or indeed Safranin(nuclear) with the excellent fast-green as the cellulosic and cytoplasmic counterstain.....
The basic method is, nuclear-stain first, remove excess with alcohol or water or a mixture of both, to isolate (differentiate) the nuclear-stain to the nuclei ideally. Then apply the counterstain to 'stain the rest' as it were, but apply this as an 'additive' stain - i.e. stain, examine, stain some more, examine etc until you have the desired figure....
A huge subject - you may try each stain on it's own first to see how it performs and is used, before serious attempts at the bi-stains.
My advice would be to forget the EY - I rarely find it satisfactory.
Use the MB as a nice general-stain and buy qalcian-blue, safranin and fast-green, in powder form, from e-bay. They're all very cheap to buy and work very well (and permanently) together..... Remember however that the above is in the context of Botanical staining.....
Search internet for safranin+fast-green staining protocol, and safranin+alcian-blue protocol - in fact if you search this forum I very likely have several posts re this with images that will help you a lot....
Good luck.
John B