Thin laser light sheet microscope for microbial oceanography
Thin laser light sheet microscope for microbial oceanography
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DOI：
10.1364/oe.10.000145
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发表时间：
2002-01-28
期刊：
影响因子：
3.8
通讯作者：
Azam, F
中科院分区：
文献类型：
--
作者：
Fuchs, E;Jaffe, JS;Azam, F
尽管需求不断增长，但海洋学家受到现有技术限制，无法观察自然环境中的水生微生物。为了提供一个简单和易于实施的解决方案，这样的研究，一个新的薄光片显微镜（TLSM）已经开发出来。TLSM利用了一个定义明确的激光片，在1 mm x 1 mm的视场内具有窄（23微米）的轴向尺寸。这个光片被精确地定位在显微镜的物镜透镜的景深内。因此，该技术利用传统的显微镜光学器件，但取代照明系统。TLSM的优点有两个方面：首先，它只在需要激发的地方聚集光，从而最大限度地提高照明光源的效率。其次，TLSM最大化图像清晰度，同时最小化背景噪声水平。不位于物镜景深内的粒子不会被照亮，因此不会造成失焦图像。来自使用SYBR绿色I荧光染色以定位单个细菌的原型系统的图像被报道。细菌在一个相对较大的和未受干扰的体积为4毫升，其中包含天然海水。TLSM可用于淡水细菌的研究，无需修改。显微镜允许在微观尺度上观察相互作用，并有可能深入了解微生物如何构建远洋生态系统。(C)2002年美国光学学会。
Despite a growing need, oceanographers are limited by existing technological constrains and are unable to observe aquatic microbes in their natural setting. In order to provide a simple and easy to implement solution for such studies, a new Thin Light Sheet Microscope (TLSM) has been developed. The TLSM utilizes a well-defined sheet of laser light, which has a narrow (23 micron) axial dimension over a 1 mm x 1 mm field of view. This light sheet is positioned precisely within the depth of field of the microscope's objective lens. The technique thus utilizes conventional microscope optics but replaces the illumination system. The advantages of the TLSM are two-fold: First, it concentrates light only where excitation is needed, thus maximizing the efficiency of the illumination source. Secondly, the TLSM maximizes image sharpness while at the same time minimizing the level of background noise. Particles that are not located within the objective's depth of field are not illuminated and therefore do not contribute to an out-of-focus image. Images from a prototype system that used SYBR Green I fluorescence stain in order to localize single bacteria are reported. The bacteria were in a relatively large and undisturbed volume of 4ml, which contained natural seawater. The TLSM can be used for fresh water studies of bacteria with no modification. The microscope permits the observation of interactions at the microscale and has potential to yield insights into how microbes structure pelagic ecosystems. (C) 2002 Optical Society of America.