| Automated Tracking of Root for Confocal Time-lapse Imaging of Cellular Processes
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Author:
Date:
2017-04-20
[Abstract] Here we describe a protocol that enables to automatically perform time-lapse imaging of growing root tips for several hours. Plants roots expressing fluorescent proteins or stained with dyes are imaged while they grow using automatic movement of the microscope stage that compensates for root growth and allows to follow a given region of the root over time. The protocol makes possible the image acquisition of multiple growing root tips, therefore increasing the number of recorded mitotic events in a given experiment. The protocol also allows the visualization of more than one fluorescent protein or dye simultaneously, using multiple channel acquisition. We particularly focus on imaging of cytokinesis in Arabidopsis root tip meristem, but this protocol is also suitable to follow ...
[摘要] 在这里，我们描述一个协议，可以自动执行增长根尖的延时成像几个小时。表达荧光蛋白或用染料染色的植物根据显影阶段的自动移动而成像，其补偿根生长，并允许随着时间跟随根部的给定区域。该协议使图像采集多个生长根尖成为可能，因此增加了给定实验中记录的有丝分裂事件的数量。该协议还允许多个荧光蛋白或染料同时显示，使用多通道采集。我们特别关注在拟南芥根尖分生组织中细胞分裂的成像，但是这种方案也适用于在各种植物物种中追随根毛发生长，花粉管生长和其他根部区域。也可以修改为自动跟踪非植物结构的根尖增长。
细胞因子是细胞分裂的最后一步，当母细胞细胞质在两个子细胞之间分配（Lipka等人，2015）时。在植物中，通过分裂平面中的细胞板的离心扩张实现，其最终成为经历有丝分裂的细胞之间的新合成的细胞壁（Buschmann和Zachgo，2016;Müller和Jürgens，2016）。植物细胞嵌入僵硬的细胞壁，不能迁移。因此细胞分裂与伸长的取向对于器官形态发生至关重要。根分生组织是研究细胞分裂的一个很好的模型，因为它们容易适用于显微技术，而不需要解剖。然而，进行细胞分裂的根长度增长，因此需要随时间手动调整观察场。该协议允许容易地延迟细胞分裂成像和其他细胞过程。
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| Stress Tolerance Assay at the Seed Germination Stage for Tobacco
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Author:
Date:
2015-04-05
[Abstract] Stress tolerance of plants is a complex phenomenon that depends on the inter-related action of several morphological, physiological and biochemical parameters. Although stress affects normal physiological growth of a plant irrespective of its developmental stage, seed germination and seed setting are considered to be the most sensitive two. Therefore, to evaluate the stress tolerance potential of a particular plant species or variety, rate of seed germination in presence of stress is an important agronomic trait. This will provide a clear indication about the stress tolerance potential with minimum instrumentation facilities. The method is very simple, effective and highly reproducible that would provide quick and reliable results to the researchers.
[摘要] 植物的胁迫耐受性是一种复杂的现象，取决于几个形态，生理和生化参数的相互作用。 虽然压力影响植物的正常生理生长，而不管其发育阶段，种子萌发和种子设置被认为是最敏感的两个。 因此，为了评价特定植物物种或品种的胁迫耐受性潜力，在胁迫存在下种子萌发的速率是重要的农艺性状。 这将提供关于最小仪器设施的应力耐受潜力的明确指示。 该方法非常简单，有效和高度可重现，将为研究人员提供快速和可靠的结果。
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