Abstract：
Cells are dynamic machines that continuously change their architecture to adapt and respond to extracellular and intracellular stimuli. Deciphering dynamic processes with nanometer-scale resolution inside cells is critical for mechanistic understanding. Here, we present a protocol that enables the in situ study of dynamic changes in intracellular structures under close-to-native conditions at high spatiotemporal resolution. Importantly, the cells are grown, transported, and imaged in a chamber in which environmental conditions such as temperature and gas (e.g., carbon dioxide or oxygen) concentration can be controlled. We demonstrate this protocol to quantify ultrastructural changes that occur during the cell cycle of cultured mammalian cells. The environment control system opens up the possibility of applying this method to primary cells, tissues, and organoids by adjusting environmental conditions.
摘要：
细胞是动态机器，不断改变其结构以适应和响应细胞外和细胞内刺激。在细胞内部以纳米级分辨率解密动态过程对于机械理解至关重要。这里,我们提出了一个方案，该方案能够在接近天然条件下以高时空分辨率对细胞内结构的动态变化进行原位研究。重要的是,细胞生长，运输,并在室内成像，其中环境条件如温度和气体(例如，二氧化碳或氧气）浓度可以控制。我们证明了该方案来量化在培养的哺乳动物细胞的细胞周期期间发生的超微结构变化。环境控制系统开辟了将这种方法应用于初级细胞的可能性，组织，和类器官通过调节环境条件。
