Special tissue dyeing method

Collagen fiber is the main component of connective tissue in the human body and is distributed in various parts of the body. The main component of collagen fibers is collagen. It is white when fresh, so it is generally called white fiber. It was dyed reddish in HE staining. Its nature is to have certain toughness and tightness, so it can resist a certain pulling force without tearing or breaking. They often aggregate into bundles of varying thickness and are wavy. Collagen fibers are composed of many fine collagen fibers. The collagen fibrils are composed of finer fibrils under electron microscopy. According to electron microscopy studies conducted by Kramen and Little, collagen fibers are unique and have a cross-grain (recycle period of 650?) fibrils. These fibers can be deformed when there is a lesion, and the collagen fibers are often necrotic. The collagen fibers can swell in the dilute acid and become a gelatinous substance. They can be dyed in shades of pink by acid dyes, which are often indistinguishable from amyloids. Because of their weaker refractive power, it is difficult to distinguish from fibers under an optical microscope. The collagen fiber has a molecular length of about 3000 Ã…, a diameter of about 14 Ã…, and a molecular weight of 300,000. (1) Composition of collagen fibers: 1. Intracellular synthesis of pre-collagen molecules, and fibroblasts take up amino acids required for synthesis of proteins, including proline, lysine and glycine. The pre-alpha-polypeptide chain is synthesized on the ribosome of the rough endoplasmic reticulum according to the base sequence of the specific gelatinous mRNA. The latter enters into the rough endoplasmic reticulum cavity while synthesizing, and under the action of hydroxylase, the hydroxylation of proline and lysine in the peptide chain is hydroxylated, and the three pre-alpha-polypeptide chains are intertwined. A rope-like procollagen molecule. The procollagen molecules in the dissolved state are unentangled at both ends and have a spherical configuration. They are secreted outside the cell after the rough endoplasmic reticulum or transferred to the Golgi complex to add a glycosyl group. 2. Extracellular polymerization of thick collagen molecules. The extracellular procollagen molecule, under the action of the endopeptidase, cuts off the spherical conformation of the two ends of the molecule to form a thick collagen molecule, which is about 1.5 nm thick and about 300 nm long. The thick collagen molecules are arranged in parallel to form a polymer. Collagen fibers. During the polymerization, the adjacent molecules which are flat with each other are shifted by 1/4 molecule length, and the molecules of the same row are kept at a certain distance from each other, and are polymerized into bundles, thereby forming collagen fibrils having a 64 nm periodic grain. During polymerization, intramolecular, intermolecular chemical genes condense and crosslink to increase the stability of thick fibers. A number of collagen fibrils are bonded to a collagen fiber such as a thick fine by a glycoprotein. Neutla-soluble collagen â†’ aldose, acetal lipid / myristic acid, lipid â†’ insoluble collagen (2) collagen fiber display: collagen fiber was stained by HE staining Dyeed in pink, in addition, it can also be dyed with some anionic dyes, such as greenish green, can be dyed blue with toluidine blue, dyed in reticular fibers If it is not treated, it can be dyed brownish yellow. Commonly used special staining methods are Van Gieson.Masson and Mallary. In immunocytochemical staining, collagen fibers often cause some non-specific staining due to excessive negative charge, and special attention should be paid. 1.Van Gieson (VG) staining method I. Preparation of reagent: weigert's hematoxylin A solution: hematoxylin 1g (haematoxylin) anhydrous alcohol 100mlB solution: 30% ferric chloride 4ml (ferric chloride) distilled water 95ml hydrochloric acid 1mlII. Van Gieson's solution A: Acid fuchsin distilled water 100mlB solution: picric acid saturated aqueous solution, (1.22%) (picric acid) Note: 1Weigert's hematoxylin is mixed with liquid A and liquid B before use. It will be used up within a few hours, and the maximum length may not exceed one day. 2 After hematoxylin is formulated, it can be matured after more than 20 days, and must be prepared in advance. 3 The liquid can resist weak acid dyeing liquid, and the effect of the dyed nuclei on the acid dyeing liquid is not easy to be removed. If it is not particularly deeply dyed, it does not need to be differentiated. 4Van Gieson's solution takes 9ml of B solution, and 1ml of A solution is added, and it can be used. Method of operation: (The various methods that follow, if not specified, are carried out at room temperature)

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