Rat submandibular and sublingual gland fragments 2 mm thick were immersed in 0.5% (0.025 W ) cyanuric chloride in anhydrous methanol containing 1% (0.1 M) N-methyl morpholine at 23-27 C. After 18-24 hr, the tissues were washed in methanol, cleared in xylene, embedded in paraffin and sectioned at 3-8 p. These tissues were compared with those fixed in: (1) 10% aqueous cold neutral buffered formalin, (2) anhydrous cold methanol,(3) 0.5% (0.014 M) cetylpyridinium chloride in 10% neutral buffered aqueous formalin, and (4) 0.4% (0.015 M) 5-aminoamidine in 50% aqueous ethanol. Tissues fixed in cyanuric chloride were well preserved with little loss or alteration of tissue components, particularly of epithelial mucins and serous components, in contrast to the p r preservation seen with the other fixatives. Tissues were examined by phasecontrast or following staining with Alcian blue 8GX and colloidal iron methods for complex acidic carbohydrates, and, the periodic acid8chiff method for carbohydrates rich in vicinal hydroxyl groups. Dyes such as orange G, aniline blue and phosphotungstic acid-hematoxylin, which do not color or are not specific for carbohydrates, were also used to reveal structural differences resulting from the various methods of fixation. The cyanuration technique is simple to perform, reproducible and safe.
Cyanuric chloride (2,4,6‐trichloro‐s‐triazine) and dichloro‐s‐triazinyl Procion M (Imperial Chemical) dyes and colorless compounds covalently bond or covalently cross‐link with H‐active functional groups in tissues including enamel, dentin, and bone. By cross‐linking, chloro‐s‐triazines make irreversibly insoluble in situ various exogenous or endogenous components, which are lost or artifactually diffused by other methods of fixation. Cyanuric chloride (0.5%) and N‐methyl morpholine (1%) in anhydrous methanol provide an excellent cytologic fixative, with N‐methyl morpholine or other suitable tertiary amines serving as “quaternizing” catalysts and HCl acceptors.Highly permeable Procion M dyes were used topically as freshly prepared 5–10% aqueous solutions; 2% solutions in 5% dextrose were also given intravenously at a dosage level of 200 mgm/Kgm. Since topically applied dyes stain irreversibly, they can be employed as markers for the study of various pathophysiologic processes associated with the tooth and periodontal apparatus; that is, for the study of growth and development. Given intravenously, these dyes permanently mark incremental lines and zones of growth in tooth and bone; the markers persist during processing and decalcification of the specimens. Vinyl‐sulfonyl Remazol (American Hoechst) and other, classes of reactive dyes were also used as permanent markers.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.