Abstract：
As live imaging plays an increasingly critical role in cell biology research, the desire to label and track individual protein molecules in vivo has been growing. To address this, in this protocol we describe steps for sparse labeling using two different HaloTag ligand dyes in C. elegans. This labeling approach is simple, is non-invasive, and preserves the view of the bulk protein population. We further describe how to carry out single-particle tracking experiments and extract information about particle diffusion behavior. For complete details on the use and execution of this protocol, please refer to Chang and Dickinson (2022).1.
摘要：
随着实时成像在细胞生物学研究中起着越来越重要的作用，在体内标记和跟踪单个蛋白质分子的愿望一直在增长。为了解决这个问题，在这个协议中，我们描述了在C.elegans中使用两种不同的HaloTag配体染料进行稀疏标记的步骤。这种标签方法很简单，是非侵入性的，并保留了大量蛋白质种群的观点。我们进一步描述了如何进行单粒子跟踪实验并提取有关粒子扩散行为的信息。有关此协议的使用和执行的完整详细信息，请参阅Chang和Dickinson（2022）。
