A Simple Chamber for Long-term Confocal Imaging of Root and Hypocotyl Development.
A Simple Chamber for Long-term Confocal Imaging of Root and Hypocotyl Development.
复制标题
DOI：
10.3791/55331
复制
发表时间：
2017-05-17
期刊：
影响因子：
--
通讯作者：
Moore I
中科院分区：
文献类型：
--
作者：
Kirchhelle C;Moore I
植物发育的几个方面，如侧根形态发生，发生在几天的时间跨度内。为了研究潜在的细胞和亚细胞过程，需要保持生理条件的高分辨率延时显微镜策略。植物组织必须有足够的营养和水供应，持续的气体交换，但当淹没和固定在盖下时，它们特别容易缺氧。一种已被成功采用的策略是使用灌注系统来维持氧气和营养物质的持续供应。然而，这种安排可能是复杂的，繁琐的，并需要专门的设备。这里提出的是一个简单的成像系统的替代策略，使用全氟萘烷作为浸没介质。该系统易于设置，需要最少的设备，并且很容易安装在显微镜台上，允许多个成像室平行设置和成像。在这个系统中，侧根的生长速度与标准条件下在琼脂板上的生长速度在前两天是难以区分的，侧根的生长速度至少在另一天以降低的速度继续。植物组织通过琼脂板提供营养，琼脂板也可用于管理一系列药理学化合物。该系统的建立是为了监测侧根的发育，但很容易适用于其他植物器官的成像，如下胚轴和初生根。
Several aspects of plant development, such as lateral root morphogenesis, occur on time spans of several days. To study underlying cellular and subcellular processes, high resolution time-lapse microscopy strategies that preserve physiological conditions are required. Plant tissues must have adequate nutrient and water supply with sustained gaseous exchange but, when submerged and immobilized under a coverslip, they are particularly susceptible to anoxia. One strategy that has been successfully employed is the use of a perfusion system to maintain a constant supply of oxygen and nutrients. However, such arrangements can be complicated, cumbersome, and require specialized equipment. Presented here is an alternative strategy for a simple imaging system using perfluorodecalin as an immersion medium. This system is easy to set up, requires minimal equipment, and is easily mounted on a microscope stage, allowing several imaging chambers to be set up and imaged in parallel. In this system, lateral root growth rates are indistinguishable from growth rates under standard conditions on agar plates for the first two days, and lateral root growth continues at reduced rates for at least another day. Plant tissues are supplied with nutrients via an agar slab that can be used also to administer a range of pharmacological compounds. The system was established to monitor lateral root development but is readily adaptable to image other plant organs such as hypocotyls and primary roots.
登录
查看更多内容
影响因子：
3.7
作者：
Sena G;Frentz Z;Birnbaum KD;Leibler S
通讯作者：
Leibler S
影响因子：
9.4
作者：
Littlejohn, George R.;Gouveia, Joao D.;Love, John
通讯作者：
Love, John
影响因子：
48
作者：
Busch, Wolfgang;Moore, Brad T.;Martsberger, Bradley;Mace, Daniel L.;Twigg, Richard W.;Jung, Jee;Pruteanu-Malinici, Iulian;Kennedy, Scott J.;Fricke, Gregory K.;Clark, Robert L.;Ohler, Uwe;Benfey, Philip N.
通讯作者：
Benfey, Philip N.
影响因子：
7.4
作者：
Beemster, GTS;Baskin, TI
通讯作者：
Baskin, TI
影响因子：
16.6
作者：
通讯作者：
--