摘 要：为高效便捷地观察禾本科植物水稻茎秆的显微结构和木质素分布，本研究采用自然干燥-冰冻切片法，成功获取薄组织切片，并结合植物自发荧光特性，运用共聚焦显微技术观察水稻茎秆的显微结构。此技术方法主要是改进了水稻茎秆冰冻切片技术，将水稻茎秆自然干燥一段时间后直接包埋、切片，可选择番红染色，也可不进行染色处理，制备出较为完整的显微结构切片。随后，利用紫外光、蓝光以及绿光激发水稻茎秆的自发荧光，进行显微成像并拍照。研究结果表明：使用改进后的冰冻切片技术可制作出厚度为1 μm且细胞结构基本完整的组织切片；使用激光扫描共聚焦显微镜收集绿光激发所产生的红色荧光信号，可快速、清晰地观察水稻茎秆切片的细胞显微结构和木质素分布情况。本研究为禾本科植物组织结构研究与生理功能探索等提供了参考。
关键词：水稻茎秆；冰冻切片技术；共聚焦显微技术；细胞显微结构；木质素分布
中图分类号：Q944.6                        文献标志码：A                    DOI：10.3969/j.issn.1007-7146.2025.04.003

Abstract: In order to observe the microscopic structure and lignin distribution of rice stalks efficiently and conveniently, this study adopted the natural drying-cryo-sectioning method and successfully obtained thin tissue sections. By combining the autofluorescence properties of plants and using confocal microscopy technique, the microscopic structure of rice stalks was observed. This technical method mainly improved the cryosectioning technique for rice stalks. After rice stalks were naturally dried for a certain period of time, they were directly embedded and sectioned. Sections can be prepared with or without safranin staining to obtain microscopic sections with relatively intact structures. Subsequently, ultraviolet light, blue light, and green light were used to excite the autofluorescence of rice stalks, followed by microscopic imaging and photography. The research results showed that the improved cryosectioning technique could produce tissue sections with a thickness of 1 μm and nearly intact cell structures. By using a laser scanning confocal microscope to collect red fluorescence signals excited by green light, the microscopic structure and lignin distribution of rice stalk sections can be observed quickly and clearly. This study provides references for the research on the tissue structures and the exploration of the physiological functions in Poaceae plants.
Key words: rice stalk; cryosection technique; confocal microscopy technique; cellular microstructure; lignin distribution
（Acta Laser Biology Sinica, 2025, 34(4): 304-311）