Kantama Laksana, Wijnker Erik, de Jong Hans
Faculty of Liberal Arts and Science, Kasetsart University, Kamphaeng Saen Campus, Kamphaeng Saen, 73140, Nakhon Pathom, Thailand.
Laboratory of Genetics, Wageningen University and Research, Droevendaalsesteeg 1, 6708 PB, Wageningen, The Netherlands.
Methods Mol Biol. 2017;1669:141-158. doi: 10.1007/978-1-4939-7286-9_12.
High-quality chromosome images of mitotic and meiotic cell divisions in plant tissues are inextricably connected with the technical control of cell spread preparations. Superb chromosome slides are the best for studying chromosome morphology and making karyotypes; they also are the best start for a successful fluorescent in situ hybridization experiment. In this study, we describe the essentials for fixation, enzymatic digestion, squash, spread, and dropping protocols for clean and well-differentiated nuclei and chromosome complements. In addition, we focus on the use of standard whole image processing for best sharpness, brightness and contrast adjustments, differentiation of heterochromatin/euchromatin, and high dynamic range imaging of big chromosomes. We also explain how to combine transparent layers or spot channels of different fluorescent images for making publication quality, full color photos.
植物组织有丝分裂和减数分裂细胞分裂的高质量染色体图像与细胞铺展制片的技术控制紧密相连。优质的染色体玻片最适合用于研究染色体形态和制作核型；它们也是成功进行荧光原位杂交实验的最佳开端。在本研究中，我们描述了用于获得清晰且分化良好的细胞核及染色体组型的固定、酶解、压片、铺展和滴片方案的要点。此外，我们着重介绍了使用标准的全图像处理来进行最佳的清晰度、亮度和对比度调整、异染色质/常染色质的区分以及大染色体的高动态范围成像。我们还解释了如何将不同荧光图像的透明层或斑点通道进行组合，以制作出具有出版质量的全彩照片。
