Stelly D M, Peloquin S J, Palmer R G, Crane C F
Stain Technol. 1984 May;59(3):155-61. doi: 10.3109/10520298409113849.
Nondissected ovaries of tuber-bearing Solanum sp. were stained with Mayer's hemalum, a positive stain for chromatin and nucleoli, and then optically cleared with methyl salicylate, a clearing agent. Clarity, resolution and contrast within the ovules dissected from ovaries were comparable to those of sectioned, paraffin embedded ovaries. Contrast within ovules greatly exceeded that of unstained and nonspecifically stained clearings, and eliminated the need of special optics, i.e., Nomarski interference-contrast optics, for optimal viewing and photography. Much less time and labor were required than for embedded specimens. Usefulness of the technique for cytogenetic and cytological research was verified by analyzing meiosis and other features of megasporogenesis and megagametogenesis in normal, and in two meiotic mutants, of Solanum. The results illustrate the usefulness of combined Mayer's hemalum staining and methyl salicylate clearing, and suggest additional stain-clearing agent combinations have potential for cytological and cytogenetic research. Preliminary results with other species suggest the technique may also be useful for classroom instruction.
对块茎类茄属植物未解剖的卵巢用梅耶苏木精染色（一种对染色质和核仁呈阳性的染色剂），然后用冬青油（一种透明剂）进行光学透明处理。从卵巢中解剖出的胚珠内的清晰度、分辨率和对比度与切片、石蜡包埋的卵巢相当。胚珠内的对比度大大超过未染色和非特异性染色透明处理后的对比度，并且无需特殊光学设备（即诺马斯基干涉对比光学设备）即可实现最佳观察和拍摄。与包埋标本相比，所需的时间和劳动力要少得多。通过分析茄属正常植株以及两个减数分裂突变体的减数分裂以及大孢子发生和雌配子体发生的其他特征，验证了该技术在细胞遗传学和细胞学研究中的实用性。结果说明了梅耶苏木精染色和冬青油透明处理相结合的有用性，并表明其他染色剂 - 透明剂组合在细胞学和细胞遗传学研究中具有潜力。对其他物种的初步结果表明，该技术也可能对课堂教学有用。
