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One aspect of Caenorhabditis elegans that makes it a highly valuable model organism is the ease of use of in vivo genetic reporters, facilitated by its transparent cuticle and highly tractable genetics. Despite the rapid advancement of these technologies, worms must be paralyzed for most imaging applications, and few investigations have characterized the impacts of common chemical anesthetic methods on the parameters measured, in particular biochemical measurements such as cellular energetics and redox tone. Using two dynamic reporters, QUEEN-2m for relative ATP levels and reduction-oxidation sensitive GFP (roGFP) for redox tone, we assess the impact of commonly used chemical paralytics. We report that no chemical anesthetic is entirely effective at doses required for full paralysis without altering redox tone or ATP levels, and that anesthetic use alters the detected outcome of rotenone exposure on relative ATP levels and redox tone. We also assess the use of cold shock, commonly used in combination with physical restraint methods, and find that cold shock does not alter either ATP levels or redox tone. In addition to informing which paralytics are most appropriate for research in these topics, we highlight the need for tailoring the use of anesthetics to different endpoints and experimental questions. Further, we reinforce the need for developing less disruptive paralytic methods for optimal imaging of dynamic in vivo reporters.

秀丽隐杆线虫（Caenorhabditis elegans）之所以成为极具价值的模式生物，核心优势之一在于体内遗传报告基因的操作便捷性——这得益于其透明的角质层与高度易操作的遗传学体系。尽管相关技术已取得飞速进展，但绝大多数成像实验仍需对蠕虫进行麻痹处理，而目前针对常用化学麻醉方法对实验测量参数（尤其是细胞能量代谢与氧化还原状态这类生化指标）的影响，相关研究仍较为匮乏。本研究采用两种动态报告基因：用于检测相对ATP水平的QUEEN-2m，以及用于反映氧化还原状态的氧化还原敏感绿色荧光蛋白（reduction-oxidation sensitive GFP, roGFP），对常用化学麻痹剂的影响展开评估。实验结果表明，在实现完全麻痹所需的剂量下，尚无任何一种化学麻醉剂能够在不改变线虫氧化还原状态与ATP水平的前提下发挥作用；且麻醉剂的使用会改变鱼藤酮暴露后相对ATP水平与氧化还原状态的检测结果。此外，我们还对常与物理束缚方法联用的冷休克处理进行了评估，发现其并不会对线虫的ATP水平与氧化还原状态产生影响。本研究不仅为相关研究领域筛选合适的麻痹剂提供了参考依据，同时强调了需根据不同实验终点与研究问题定制化选择麻醉方案的必要性。更进一步，本研究也呼吁开发干扰性更低的麻痹方法，以实现动态体内报告基因的最优成像检测。