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环境卫生:评估我们的水世界的流式细胞术方法。

Environmental health: flow cytometric methods to assess our water world.

作者信息

Yentsch C M

机构信息

J.J. MacIsaac Flow Cytometry/Sorting Facility, Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Maine 04575.

出版信息

Methods Cell Biol. 1990;33:575-612. doi: 10.1016/s0091-679x(08)60555-5.

Abstract

Flow cytometry/cell sorting in aquatic sciences has been driven in two directions. The frontier directions are on shipboard and shore-based. On the one hand, the rapid analytical technique has been taken on shipboard to provide a real-time assessment of the particles and phytoplankton in water masses. These data also give information on the amount of vertical mixing and advection, and denote fronts between two or more water masses. There is an optical characterization (based on sizes, numbers, and pigment groups) of the individual primary producers, as well as detritus and suspended sediments. An optical-closure question is being addressed: "Does the total optical signal equal the sum of the parts?" Additionally, associations with chemical and physical oceanographic features are readily accomplished. A "census" of thousands of phytoplankton cells is obtained and can be mapped. Scientists are able to identify "who is where?" Such data are critical to understand the optical-feedback loop or the so-called photon-budget-in-the-sea, which in turn controls the rates at which growth processes occur in nature. On the other hand, an in-depth understanding is sought as to how particle size, shape, refractive index, nutritional status (nutrient and/or light limitation), growth dynamics, and cell cycle combine to control the optics (light scatter and fluorescence at the moment, and ideally absorption as well) or the photon-budget-of-the-cell. For this purpose, a shore-based facility associated with a diverse collection of phytoplankton is ideal. The development at Bigelow Laboratory of the Jane J. MacIsaac Facility is to provide services for the oceanographic community. Association and co-location with the Provasoli-Guillard Center for Culture of Marine Phytoplankton is key. Visitors are trained and given access to state-of-the-art instrumentation. Visiting investigators have available "the tropical, temperate, and polar seas" in concentrated form, as marine phytoplankton isolated worldwide and maintained as living clonal cultures. In this way, frontline cell biology questions can be addressed. The relentless exploration of standards and controls appropriate for the aquatic community must be continued. An intercalibration effort is a vital step. It is only with the widespread acceptance of particular reference materials and uniform optical filters among research groups utilizing FCM that comparable data sets describing aquatic particle distributions will be possible. For a global science, this strategy is imperative.

摘要

水生科学中的流式细胞术/细胞分选朝着两个方向发展。前沿方向是在船上和岸上。一方面,快速分析技术已应用于船上,以实时评估水体中的颗粒和浮游植物。这些数据还提供了有关垂直混合和对流程度的信息,并表明两个或更多水体之间的锋面。对单个初级生产者以及碎屑和悬浮沉积物进行了光学表征(基于大小、数量和色素组)。一个光学封闭问题正在得到解决:“总光学信号是否等于各部分信号之和?”此外,与化学和物理海洋学特征的关联很容易实现。获得了数千个浮游植物细胞的“普查”结果并可进行绘图。科学家们能够确定“谁在哪里?”此类数据对于理解光学反馈回路或所谓的海洋光子预算至关重要,而这反过来又控制着自然界中生长过程发生的速率。另一方面,人们正在深入了解颗粒大小、形状、折射率、营养状况(营养和/或光照限制)、生长动态和细胞周期如何共同控制细胞的光学特性(目前的光散射和荧光,理想情况下还有吸收)或细胞的光子预算。为此,与各种浮游植物集合相关的岸上设施是理想之选。比奇洛实验室的简·J·麦金太尔设施的发展旨在为海洋学界提供服务。与普罗瓦索利 - 吉拉德海洋浮游植物培养中心的关联和并置是关键。为参观者提供培训并让他们使用最先进的仪器设备。来访的研究人员可以以浓缩形式获取“热带、温带和极地海域”的样本,这些样本是从世界各地分离并作为活克隆培养物保存的海洋浮游植物。通过这种方式,可以解决前沿的细胞生物学问题。必须持续不懈地探索适用于水生生物群落的标准和控制方法。相互校准工作是至关重要的一步。只有在使用流式细胞术的研究小组中广泛接受特定的参考材料和统一的光学滤光片,才有可能获得描述水生颗粒分布的可比数据集。对于一门全球科学而言,这一策略至关重要。

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