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UND 363 Mass. Tri.
UND 363 Masson Trichrome
| Question | Answer |
|---|---|
| Purpose for Masson Trichrome | distinguish collage from muscle (helps ID diseases with increased fibrotic changes) often bowel, liver and kidney Bx's |
| What is used initially | bouins used as mordant to link dye to tissue |
| what is the nuclei stained with | Weigerts (iron hematox). Used early in stain because of the subsequent acid solutions |
| what do the 3 dyes stain | muscle, collagen, fibrin, and RBC |
| what does the biebrich scarlet acid fuschin stain | stains acidophilic elements (cytoplasm, muscle, collagen) |
| what is the purpose of the phosphomolybdic/phosphotungstic adic? | decolorizes/diffuse out collagen fibers leaving the muscle cells red |
| what is the purpose of the aniline blue soln' | links with collagen (gives blue color) |
| what does acetic acid do in this stain | differentiation (delicate color shades and transparency) |
| typical fixatives | bouins preferred, but 10%NBF OK |
| Control | none needed (most tissue has built in control), but uterus, sm. int. appendix, or fall. tube can be used |
| microns cut at | 4-5 |
| what are the 6 reagents used in this stain | picric acid (optional pretreatment), bouins (mordant), weigert hematox (stains nuclei and is acid resistant), biebrich scarlet, phosphotung/molybdic acid, aniline blue, acetic acid |
| give facts about phosphotungstic/phosphomolybdic acid | numerous acid groups to link decolorized collagen to aniline blue, ph of PTPM increases selective collagen staining and aids in removal of biebrich scarlet, must be fresh, if inadequate stain length the aniline blue will not stain. |
| results | nuclei - black, cytoplasm/keratin/muscle - red, collagen and mucous - blue |
| give order of reagents | bouins, weigerts, biebrich scarlet, PTPM, aniline blue, acetic acid |
| what can cause decreased red staining | aged solutions, overused solutions ( fix is to make new and restain) |
| what can cause faded/cecreased blue | overdiff'd with acetic acid, pathologic conditions where collagen is altered (ie burns) |
| what can affect the blue staining of collagen | increasing or decreasing the incubation time in aniline blue |
| what are the 2 methods for masson trichrome (and the advantage) | conventional and microwave (m'wave accelerates steps and uses gills hematox III) |
| what can be used as a counter stain instead of aniline blue | lt green (is better if there is an abundance of collagen) |
| why should caution be used with picric acid | explosive with less than 10% h20 (use jar in a jar method) |
| Principle of Masson Trichrome | 3 dyes, b. scarlet stains first, phosphotung/molybd used second and diffuse out b. scarlet from collagen but not cytoplasm and link the collagen with w/ aniline blue. Acetic acid used to differentiate. |
Created by:
mustangvxd
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