MICROSPECTROPHOTOMETRIC ANALYSIS OF METACHROMATIC STAINING OF NUCLEIC ACIDS
MICROSPECTROPHOTOMETRIC ANALYSIS OF METACHROMATIC STAINING OF NUCLEIC ACIDS
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DOI：
10.1086/physzool.25.4.30152126
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发表时间：
1952-01-01
期刊：
影响因子：
--
通讯作者：
HIMES, MH
中科院分区：
文献类型：
--
作者：
FLAX, MH;HIMES, MH
天青B是一种异染性染料，当在pH 4下使用时，特异性地在卡诺伊固定的植物和动物组织中染色核酸和酸性多糖。测定了染料与核酸结合后在组织中的吸收曲线。用显微分光光度计650 m/x处的正色吸收峰（[alpha] -峰）始终存在，而590 m[mu]处的2d，[beta]-峰和545 m[mu]附近的3d，[mu]-峰也可能出现，最后一个峰导致在某些染色条件下观察到的异染性颜色。[alpha]_和[beta]-峰与浓缩物的峰相同。水溶液而《易经》中的《易经》，则是一个经典。与溶液中与某些显色剂结合的染料的[mu]峰相同的位置。这3个峰的重复出现支持了基于溶液中染料聚合的异染性理论的扩展。来解释组织中的异染现象改变染色条件的效果似乎可以用这一理论满意地解释。增加染料浓度。pH和温度。(up至60[degree]C）的染色溶液。增加核酸的异染性。这可能是由于amt的增加。染料结合，这大概会增加染料聚合的可能性。在染料浓度范围内。0.2-0.4毫克/ CC.在40 ℃时，RNA染异染性（紫色），而DNA相对地染正染性（蓝绿色），这使其成为用于组织切片中核酸的差异定位的有用技术。这种差异的程度的异染性的RNA和DNA的分子结构方面的影响进行了讨论。
Azure B, a metachromatic dye, when used at pH 4, stains nucleic acids and acid polysaccharides specifically in Carnoy-fixed plant and animal tissues. Absorption curves of the dye bound to nucleic acids in tissues were detd. with a micro-spectrophotometer. The orthochromatic absorption peak ([alpha] -peak) at 650 m/x is always present, while a 2d, [beta]-peak at 590 m[mu] and a 3d, [mu]-peak near 545 m[mu], may also occur, the last giving rise to the metachromatic color observed under some staining conditions. The [alpha]_ and [beta]-peaks are identical with those of a concd. aqueous soln. of the dye, while the [mu]-peak is in approx. the same position as the [mu]-peak of the dye combined with certain chromotropes in soln. The repeated occurrence of these 3 peaks supports an extension of the theory of metachromasy based on dye polymerization in soln. to explain metachromasy in tissues. The effect of alterations of the staining conditions appears to be satisfactorily interpreted by this theory. Increasing dye concn. pH, and temp. (up to 60[degree]C) of the staining soln. increases metachromasy of nucleic acid. This may result from the increase in the amt. of dye bound, which presumably would increase the probability of dye polymerization. Within the range of dye concns. of 0.2-0.4 mg./cc. at 40[degree]C, RNA stains metachromatical-ly (purple), while DNA stains relatively orthochromatic ally (blue-green), making this a useful technic for the differential localization of nucleic acids in tissue sections. The implications of this difference in the degree of metachromasy of RNA and DNA in terms of their molecular structure are discussed.