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冬虫夏草和蛹虫草作为重要的药用真菌,得到广泛的重视,迄今已有大量的研究报道。然而,作为细胞学研究的基础处理方法,其菌丝隔膜和细胞核染色却缺乏必要的研究。荧光染色是一种程序简便、快速灵敏的染色方法。本研究选用DAPI、PI、Calcoflour White和刚果红等4种染料,对冬虫夏草、蛹虫草菌丝的隔膜和细胞核进行单独与组合染色实验,通过显微观察比较得出较好的染色方法。结果表明DAPI对冬虫夏草和蛹虫草菌丝的细胞核染色效果都较好,而Calcoflour White对两者的细胞壁染色效果较好且隔膜清晰。DAPI与Calcoflour White两者进行冬虫夏草菌丝组合染色的效果为最佳,但在蛹虫草菌丝染色中效果不太稳定。对蛹虫草菌丝较好的组合染色是DAPI与刚果红的组合,但其染色结果需要在激光共聚焦显微镜下观察。
As an important medicinal fungus, Cordyceps sinensis and Cordyceps militaris have received a great deal of attention. So far, a large number of studies have been reported. However, as a basic treatment method for cytology, its mycelial septum and nuclear staining lack the necessary research. Fluorescent staining is a simple, fast and sensitive staining method. In this study, DAPI, PI, Calcoflour White and Congo red were used to stain the membrane and nucleus of Cordyceps sinensis and Cordyceps militaris individually and in combination, and the better dyeing method was obtained by microscopic observation. The results showed that DAPI staining of Cordyceps sinensis and Cordyceps militaris nuclei were better, and Calcoflour White staining of both the cell wall better and clear membrane. The combination of DAPI and Calcoflour White for the best combination of Cordyceps sinensis mycelium, but the effect of Cordyceps militaris is not stable. The preferred combination of DAPI and Congo red for Cordyceps militaris mycelial staining, but the staining results need to be observed under a laser confocal microscope.