不同扫描电镜制样方法观察棉花叶片气孔

Different SEM Sample Preparation Methods to Observe the Stomata of Cotton Leaves

扫描电子显微镜是研究棉花气孔发育及形态建成的重要手段。为了分析常规扫描电子显微镜、环境扫描电子显微镜、冷冻扫描电子显微镜、涂撕法扫描电子显微镜4种方法在棉花气孔观察应用上的优劣,本文通过对棉花叶片取材、制样、镜检,横向对比了这4种方法的取样方式、制样时间和镜检结果。结果表明:常规扫描电子显微镜生物制样时间最长,样品有一定的失水,图片衬度效果最好;环境扫描电子显微镜制样时间最短,样品失水严重,图片衬度较差,样品形态与常规扫描电子显微镜生物制样方法所观察到的形态一致;冷冻扫描电子显微镜制样时间较长,样品未发现失水,图片衬度较差;涂撕法扫描电子显微镜制样时间较短,样品未发现失水,图片衬度较好,但相对于冷冻扫描电子显微镜制样方法而言,其样品呈现出反向的形态,同时由于尖端放电,荷电较多。本研究分析了不同的扫描电子显微镜制样方法适合得到怎样的棉花气孔图片数据,为棉花抗旱相关的基础和应用研究提供了一定的参考。

Scanning electron microscope(SEM) is an important way to study the development and morphogenesis of cotton stomata. Herein, advantages and disadvantages in cotton pore observation with SEM, environmental scanning electron microscope(ESEM), cryo-SEM, and print-SEM were analyzed by comparing horizontally their sampling methods, including the sample collection, preparation time and microscopy results. SEM proved to have the longest sample preparation time, some water loss in samples, but the best image contrast.ESEM took the shortest time to make samples, but had serious water loss in samples and poor image contrast. The morphology of the samples was consistent with that observed by the SEM biological sampling method. Cryo-SEM took a long time to prepare samples, and there was no water loss in the samples, but poor image contrast. The sample preparation time for print-SEM was short, with no water loss and a better image contrast and the sample presented the morphology of the reverse cryo-SEM, but the charge was more due to the tip discharge. This study revealed what kind of stomatal image data could be obtained by different SEM sampling methods, and may provide some references for research on drought-resistant basis and application of cotton.