Evaluation of preparation, staining and microscopic techniques for counting incremental lines in cementum of human teeth
Evaluation of preparation, staining and microscopic techniques for counting incremental lines in cementum of human teeth
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DOI：
10.3109/10520299609117155
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发表时间：
1996-07-01
影响因子：
1.6
通讯作者：
Bjerketvedt, D
中科院分区：
文献类型：
--
作者：
Kvaal, SI;Solheim, T;Bjerketvedt, D
牙骨质的附着发生在两个阶段，导致两种类型的层具有不同的光学和染色性质，可以通过光学显微镜观察到，窄的深色染色增量线被较宽的浅色染色牙骨质带分开。从一条线到下一条线的距离代表了许多哺乳动物牙骨质存款的年增量，并且计数这些线已被常规地用于估计动物的年龄。牙骨质中的增量线也已在人类牙齿的切片中观察到，并且本研究的目的是检查用于计数的制备和染色的许多方法。从福尔马林固定的人牙根上切下研磨或脱钙的样品，石蜡包埋或冷冻，并使用几种技术染色。采用常规光、荧光、偏振光、共聚焦激光扫描、干涉衬度、相差和扫描电子显微镜对牙骨质进行了研究。牙骨质中的增量线可以观察到在地面切片，脱钙后，在冷冻和石蜡包埋切片。甲苯胺蓝、甲酚紫、苏木精或高碘酸希夫（PAS）染色的增量线允许通过常规光学显微镜进行区分，使用荧光显微镜和绿色光激发的对比度最好，因为染色的牙骨质带，而不是增量线，在用甲酚紫、PAS或苏木精和伊红染色后发出荧光。其他显微技术的结果不令人满意。由于增量线不会被酸破坏，并且与剩余的牙骨质不同，因此它们可能具有与牙骨质不同的有机结构。使用绿色光激发的甲酚紫染色的脱钙切片可以最好地观察人牙骨质中的增量线。
Apposition of cementum occurs in phases resulting in two types of layers with different optical and staining properties that can be observed by light microscopy, Narrow, dark staining incremental lines are separated by wider bands of pale staining cementum. The distance from one line to the next represents a yearly increment deposit of cementum in many mammals, and counting these lines has been used routinely to estimate the age of the animals, Incremental lines in cementum have also been observed in sections of human teeth, and the object of the present investigation was to examine a number of methods for preparing and staining them for counting, Longitudinal and transverse sections, either ground or decalcified, were cut from formalin fixed human dental roots, paraffin embedded or frozen, and stained using several techniques. The cementum was investigated using conventional light, fluorescence, polarized light, confocal laser scanning, interference contrast, phase contrast, and scanning electron microscopy. Incremental lines in the cementum could be observed in ground sections and, following decalcification, in both frozen and paraffin embedded sections. Toluidine blue, cresyl violet, hematoxylin, or periodic acid Schiff (PAS) stained incremental lines allowing differentiation by conventional light microscopy, Contrast was best using fluorescence microscopy and excitation by green light since the stained cemental bands, but not the incremental lines, fluoresced after staining with cresyl violet, PAS or hematoxylin and eosin. The results with other microscopic techniques were unsatisfactory. Since incremental lines are not destroyed by acids and stain differently than the remaining cementum, it is likely that they possess an organic structure which differs from the cementum, Incremental lines in human dental cementum could be observed best using decalcified sections stained with cresyl violet excited by green light.