冬虫夏草、蛹虫草菌丝隔膜和细胞核荧光染色

Fluorescent staining of septa and nuclei in Ophiocordyceps sinensis and Cordyceps militaris

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 important medicinal fungi, both Ophiocordyceps sinensis and Cordyceps militaris have gained significant scientific attention in recent decades. However, very little work on mycelial staining method, which is the basis of cytological research, has been reported. Fluorescent staining is a simple and fast staining technique. In the present study, four dyes were tested, singly and combinedly, for use on Ophiocordyceps sinensis and Cordyceps militaris as possible fluorescent stains of fungal nuclei and septa. The results showed that DAPI fluorescent stained better than PI for the nuclei, and the Calcoflour White is better than Congo Red in staining the septa. In the combined stainings, DAPI and Calcoflour White stained nuclei and septa, respectively, more effectively in O. sinensis than in C. militaris, whereas the combination of DAPI and Congo Red is more suitable for the latter by using the laser confocal microscope.