A VERSATILE STAIN FOR POLLEN FUNGI, YEAST AND BACTERIA
A VERSATILE STAIN FOR POLLEN FUNGI, YEAST AND BACTERIA
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DOI：
10.3109/10520298009067890
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发表时间：
1980-01-01
期刊：
影响因子：
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通讯作者：
ALEXANDER, MP
中科院分区：
文献类型：
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作者：
ALEXANDER, MP
开发了一种通用的染色方法，用于显示花粉、真菌菌丝和孢子、细菌和酵母。该混合物按以下顺序混合制成：乙醇20毫升；1%孔雀绿95%乙醇2毫升；蒸馏水50毫升；甘油40毫升；酸性品红1%蒸馏水10毫升；苯酚5克和乳酸1-6毫升。还配制了去除沾满花粉的溶液。理想情况下，败育的花粉粒染成绿色，未败育的花粉粒染成深红色。真菌菌丝和孢子呈蓝紫色，寄主组织呈绿色。真菌、细菌和酵母菌被染成紫色到红色。混合染液中乳酸的浓度对花粉的区别染色起着重要作用。对于真菌、细菌和酵母的染色，着色剂必须是酸性的，但除了细菌外，其浓度并不是关键。对于花粉，染色可以在玻片上的一滴污渍中进行，也可以在小瓶中的几滴污渍中进行。染在瓶子里的花粉可以立即使用，也可以储存起来供以后使用。轻轻燃烧载玻片或以55.+-的温度保存可加速染色。2.学位。C培养24小时。细菌和酵母菌按常规方式固定在玻片上，然后染色。原液经久耐用，染色混合物非常稳定，安装的标本在长时间储存后不会褪色。幻灯片是半永久性的，如果盖页下面出现气泡，只要添加1-2滴污渍，就没有必要环住盖页。还讨论了在图像分析仪中使用差异染色的花粉片来自动计数和记录败育和未败育的花粉。
A versatile stain was developed for demonstrating pollen, fungal hyphae and spores, bacteria and yeasts. The mixture is made by compounding in the following order: ethanol, 20 ml; 1% malachite green in 95% ethanol, 2 ml; distilled H2O, 50 ml; glycerol, 40 ml; acid fuchsin 1% in distilled H2O, 10 ml; phenol, 5 g and lactic acid, 1-6 ml. A solution was also formulated to destain overstained pollen mounts. Ideally, aborted pollen grains are stained green and nonaborted ones crimson red. Fungal hyphae and spores take a bluish purple color and host tissues green. Fungi, bacteria and yeasts are stained purple to red. The concentration of lactic acid in the stain mixture plays an important role in the differential staining of pollen. For staining fungi, bacteria and yeasts, the stain must be acidic, but its concentration is not critical except for bacteria. In the case of pollen, staining can be done in a drop of stain on a slide or in a few drops of stain in a vial. Pollen stained in the vial can be used immediately or stored for later use. Staining is hastened by lightly flaming the slides or by storing at 55 .+-. 2.degree. C for 24 h. Bacteria and yeasts are fixed on the slide in the usual manner and then stained. The stock solution is durable, the staining mixture is very stable and the color of the mounted specimens does not fade on prolonged storage. Slides are semipermanent and it is not necessary to ring the coverslip provided 1-2 drops of stain are added if air bubbles appear below the coverslip. The use of differentially stained pollen mounts in image analyzers for automatic counting and recording of aborted and nonaborted pollen is also discussed.