Alcon Research, LLC.
Alcon Research, LLC;
J Vis Exp. 2024 Sep 20(211). doi: 10.3791/66268.
Acanthamoeba keratitis is a severe ocular infection that poses treatment challenges and can lead to blindness. Despite its ubiquity and potential contamination of contact lenses after water exposure, the natural behavior of this pathogen remains elusive. Understanding Acanthamoeba's movement patterns can inform us about how it colonizes contact lenses and contaminates the patient's cornea. It is fortunate that Acanthamoeba spp. are visible via brightfield microscopy starting at 4x magnification. Previous techniques have been developed to quantify Acanthamoeba motility in regard to cytopathic effects or under-exposure to the electric field. Here, we describe a method to track and quantify Acanthamoeba spp. motility long-term (hours to days), which is a protocol applicable to multiple amoeba strains, surfaces, and nutritional status of amoeba. This procedure is germane to determining many core motility quantifications, such as distance, speed, confinement, and directionality, which are necessary to monitor different stages of infection, proliferation, or behavioral change.
棘阿米巴角膜炎是一种严重的眼部感染，治疗具有挑战性，可导致失明。尽管棘阿米巴无处不在，并且在接触镜暴露于水后可能会被污染，但这种病原体的自然行为仍然难以捉摸。了解棘阿米巴的运动模式可以帮助我们了解它如何定植于接触镜并污染患者的角膜。幸运的是，棘阿米巴属在 4x 放大倍数的明场显微镜下即可观察到。先前已经开发出一些技术来量化与细胞病变效应或电场暴露不足有关的棘阿米巴运动。在这里，我们描述了一种跟踪和量化棘阿米巴属运动的长期（数小时到数天）方法，这是一种适用于多种阿米巴株、表面和阿米巴营养状况的方案。该程序与确定许多核心运动学定量分析有关，例如距离、速度、约束和方向性，这些定量分析对于监测感染、增殖或行为改变的不同阶段是必要的。
