Stains to start with

[charlie]

I see we need acid and basic dyes. I assume we novices need a few stains to start with?

What would you suggest for beginners? I really want a gallon of this stuff but i bet it isn't on the starter list!

http://www.ebay.com/itm/321811741166

[JimT]

Charlie, I use to only used iodine because it brings out the nucleus so well. Just recently I added methylene blue to my repertoire and it brings out the internal cell cytoplasm nicely. I would recommend those two to start with.

Maybe other more experienced stain users can add more suggestions.

JimT

[charlie]

Thanks for the reply JimT.

Looking on amazon this comes up and seems the least expensive way to buy Methylene Blue:
http://www.amazon.com/Methylene-Blue-Ge ... VE5DEQK9GT

4 oz for$5.59 and it is zinc free, chloride salt of Methylene Blue. Provided as a 2.303% water solution.

That is twice the concentration at under half the price of:
http://www.amazon.com/Ricca-Chemical-Me ... P8SWSNZ0FC

Am I missing something?

[JimT]

More than I know. I bought my bottle from a pet store fish area. Methylene Blue is also used to treat aquarium fish ick.

[travis]

More than I know. I bought my bottle from a pet store fish area. Methylene Blue is also used to treat aquarium fish ick.

Do you need to dilute the M/blue you got from the pet store?

[mrsonchus]

More than I know. I bought my bottle from a pet store fish area. Methylene Blue is also used to treat aquarium fish ick.

Do you need to dilute the M/blue you got from the pet store?

You'll need to check the concentration on the bottle and dilute as needed, I routinely use 0.05% (aq) as it's a strong stain, but for hand sections I'd probably start with 0.1% and see how the tissue reacts. Good luck.

[travis]

Unfortunately the one i bought does not have the concentration level, i'll dilute several in some syringes and see what works best.
Thanks.

[mrsonchus]

Unfortunately the one i bought does not have the concentration level, i'll dilute several in some syringes and see what works best.
Thanks.

That'll do it Travis, remember that a duration of >= about 2 mins is usually best for control of staining-effect.
The basic method is;

1) into stain (we'll assume aqueous) for time.
2) rinse with water until excess stain stops or virtually stops coming away from tissue (this isn't removing stain from tissue that has 'taken', only excess stain).

It's also possible (and easy) to differentiate when you've got the basic concentration/duration sorted. This will remove stain from different structures or cell types at different rates, leaving behind tissue that 'differentiates' by virtue of varying shades and strengths of stain retention. It's also very helpful to 'mature' Methylene blue by exposure to air, this will increase the ability of the stain to act metachromatically and again give a sort of 'one-stop' differentiation by virtue of 3 isomers that form within the stain, effectively resulting in a 'trichrome', albeit in shades of blue.
An acidic-alcohol rinse will cause differentiation by removal, without acidification (usually with a touch of acetic acid) the alcohol will not really remove any Methylene blue and may therefore be used to stop the effect of the acid when the desired effect is reached...

Good luck, there's an awful lot that can be achieved with Methylene blue, a very underrated stain in my experience.