Microfluidic Device for High-Resolution Cytoskeleton Imaging and Washout Assays in Physcomitrium (Physcomitrella) patens
Microfluidic Device for High-Resolution Cytoskeleton Imaging and Washout Assays in Physcomitrium (Physcomitrella) patens
复制标题
用于小立碗藓 (Physcomitrella) 高分辨率细胞骨架成像和冲洗测定的微流体装置
DOI：
10.1007/978-1-0716-2867-6_11
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发表时间：
2023
影响因子：
--
通讯作者：
E
中科院分区：
文献类型：
--
作者：
Yoshida;M.W.;Kozgunova;E
以高时空分辨率可视化细胞骨架动力学，可以深入了解动力学变化方式及其与多种蛋白质的相互作用，以维持细胞功能。倾斜照明荧光显微镜是一种对质膜附近的细胞事件进行成像的流行技术。在本章中，我们提供了在微流体装置中使用苔藓立碗藓（Physcomitrium patens）的原丝体和配子体细胞进行高分辨率细胞骨架成像的详细协议。这些包括聚二甲基硅氧烷（PDMS）装置的制备、苔藓细胞的培养以及短期和长期倾斜照明荧光显微镜。我们还描述了如何在活细胞成像过程中将化合物（例如微管破坏药物）引入设备并从设备中洗掉。
Visualizing cytoskeleton dynamics at high spatiotemporal resolution provides valuable insights into the way the dynamics change as well as its interactions with multiple proteins in order to maintain cellular function. Oblique illumination fluorescent microscopy is a popular technique to image cellular events localized near the plasma membrane. In this chapter, we provide detailed protocols for high-resolution cytoskeleton imaging using protonema and gametophore cells of the moss Physcomitrella (Physcomitrium patens) in the microfluidic device. These include preparation of the polydimethylsiloxane (PDMS) device, culture of moss cells, and both short- and long-term oblique illumination fluorescent microscopy. We also describe how to introduce to, and wash out from, the device chemical compounds, such as microtubule-disrupting drugs, during live-cell imaging.