A rapid and precise technique for enumeration of resting cysts of Alexandrium spp. (Dinophyceae) in natural sediments
A rapid and precise technique for enumeration of resting cysts of Alexandrium spp. (Dinophyceae) in natural sediments
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一种快速、精确的亚历山大藻休眠包囊计数技术。
DOI：
10.2216/i0031-8884-34-3-207.1
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发表时间：
1995
期刊：
影响因子：
1.6
通讯作者：
Y. Ishida
中科院分区：
文献类型：
--
作者：
M. Yamaguchi;S. Itakura;I. Imai;Y. Ishida
摘要本文介绍了一种利用荧光原色显微镜和会聚荧光显微镜对天然沉积物中鞭毛藻囊进行精确计数的新方法。用戊二醛固定实验室培养的塔玛兰Alexandrium tamarense (Lebour) Balech静息囊，并用甲醇处理。在相同的程序下，用九种荧光染料对囊肿进行染色，以找到适合计数的囊肿。四种荧光染料，吖啶黄、钙白M2R、尼罗河红和樱草碱，在高染色度和荧光强度方面提供了令人满意的结果。固定后需进行甲醇处理，以获得较高的染色度。然后检查了四种荧光染料在广岛湾沉积物中枚举自然发生的囊肿的适用性。春草碱被证明优于所有其他染料，提供更高的囊肿计数。原花素染色的囊肿在蓝光激发下表现出强烈的黄绿色荧光。
Abstract A new method is described that uses the fluorochrome primuline and epifluorescence microscopy for precise enumeration of dinoflagellate cysts in natural sediments. Alexandrium tamarense (Lebour) Balech resting cysts obtained in laboratory culture were fixed with glutaraldehyde and treated with methanol. The cysts were stained using nine fluorochromes under identical procedures to find those suitable for enumerating cysts. Four fiuorochromes, acriflavine, calcofluor white M2R, nile red and primuline, were found to provide satisfactory results in terms of high stainability and fluorescence intensity. Methanol treatment after fixation was necessary for high stainability. The four fluorochromes were then examined for their applicability to enumerate naturally occurring cysts in Hiroshima Bay sediments. Primuline proved to be superior to all other dyes examined, providing higher counts of cysts. Primuline-stained cysts exhibited an intense yellow-green fluorescence under blue-light excitation which hi...