Trichrome staining

[Naphthalene]

Hello everyone!
I have recently attempted to stain a mouse embryo trunk horizontal sections using Masson's trichrome method, using the IHCworld protocol with few modifications. Ponceau instead of fuchsine, some different timings. I skip the details..

Its a17 day mouse embryo, thats a secion through a belly area. You can see the spinal cord inside the future vertebrae (blue), the developing lung, liver (bottom right) and the outer skin with underlying mucle tissues. 8 micron section, Brunel rocker.

Blue is the collagen (connective tissue), red - other proteins like muscle myosin & skin keratin, dark-blue/black - cell nuclei. Erythrocytes - orange-red.

[gekko]

Naphthalene, excellent images with beautiful colors! I am sure a lot of expertise and hard work went into the preparation and staining, but the results speak for themselves.

[JimT]

Your industry continues to impress me. How big is a mouse embryo trunk horizontal section and where did you get it and how did you section it?

BTW, great images!

JimT

[mrsonchus]

Beautifully clear differentiation - simply superb!
Very, very nice indeed.

[Seb28]

Very nice,I like the 2 image,very clear and sharp.

[Naphthalene]

Dear gekko, mrsonchus and Seb28 thank you for your kind comments!

This is a 17-18 day old mouse embryo, it is about 7 mm in diameter.

I don't work with animals myself, I get my mouse samples as a "waste product" from our animal facility, in the current example the guys were using mouse embryonic brain tissue, while the trunks were discarded. I asked them to save these trunks for me, so I have a half-dozen of them fixed in formalin in my home fridge (to scare the parents if they are to ventrue inside my "vegetable" box at the bottom )

The paraffin embedding and sectioning were performed by standard methods. Before sectioning. the blocks were soaked in 10% ammonia for an hour, chilled with ice and immediatelly cut. Sections were stretched at 48-50C on water (<- thats the hardest part - the bone stretches ugly and gives rise to wrinkles) and mounted on gelatin coated slides.

The staining is straightforward, I simply followed the online protocol for Masson's and I did not attempt to optimize the conditions.

[Raul]

Hi Naphthalene, congratulation for the samples, they are indeed with a lot of information and also they present a sweet for the eyes. I do have a question for you, did you post-process the images with a software (like photoshop or something similar) ?