Mallory Method

MALLORY METHOD (Mallory), more correctly Mallory's methods for staining collagen fibers. First method: 1) any fixation, alcohol is best; 2) preparation of sections, frozen or after embedding in celloidin or paraffin; 3) immersion of sections for 30 seconds in a 10% aqueous solution of phosphomolybdic acid (only glass needles are used); 4) washing in water; 5) staining for 10-20 minutes in the following solution: 10% phosphomolybdic acid 10 cm³, crystalline hematoxylin 1.75 g, water 200 cm³, crystalline carbolic acid 5 g (the solution is placed in the sun for several hours before use and filtered); 6) rinsing in water; 7) dehydration in alcohol, xylene, balsam. Collagen fibers are dark blue. Dehydration is best performed with origanum oil instead of alcohol. Phosphomolybdic acid can be replaced by phosphotungstic acid with mandatory differentiation after staining and rinsing in water and in 50% alcohol. - S e c o n d Mallory method: 1) fixation in Zenker's fluid (not Zenker-formol!); 2) freezing or embedding; 3) staining for 1-3 minutes in a heavily (10-20 times) diluted with water aqueous 1% solution of acid fuchsin; 4) rinsing in water; 5) treatment of sections for 5-15 minutes in a 1% aqueous solution of phosphomolybdic acid; 6) rinse twice in pure water; 7) staining for 2 minutes or more in the mixture: Anilin-blau wasserlöslich - 0.5 g, Orange G - 2 g, oxalic acid - 2 g, water 100 cm³ (the mixture is heated to boiling and filtered upon cooling); 8) washing in water, differentiation in pure 96° alcohol and dehydration in absolute alcohol (can be replaced by essential oils - origanum, bergamot). Only pure alcohol is used; formalin or denatured alcohol is not permissible. Clearing in xylene. Balsam. Collagen fibers - dark blue; the same for amyloid, mucus, hyaline; cell nuclei, fibrin, elastin - red; protoplasm, axis cylinders of nerve fibers and neuroglia - violet; erythrocytes, myelin sheaths of nerves - yellow or yellow-red; cornified substance - bright red; muscles - fire-red. Before staining according to Mallory, it is useful to stain the nuclei for 24 hours in alum carmine (Alauncarmin). Sections from material fixed in formalin are best pre-treated in Zenker's fluid for 2-24 hours and washed in water. - The original Mallory method is currently often replaced by various modifications. Among them, the most successful is the modification proposed by M. Heidenhain (the so-called "Azan"). Here, fuchsin is replaced by azocarmine, and Mallory's mixture is modified. In some cases, it is very good to replace fuchsin with alum carmine (Alauncarmin, 24 hours). Fuchsin can also be successfully replaced with iron hematoxylin (Mislavsky). See also Neuroglia.