Abstract：
The tear film is a highly dynamic biofluid capable of reflecting pathology-associated molecular changes, not only in the ocular surface but also in other tissues and organs. Molecular analysis of this biofluid offers a non-invasive way to diagnose or monitor diseases, assess medical treatment efficacy, and identify possible biomarkers. Due to the limited sample volume, collecting tear samples requires specific skills and appropriate tools to ensure high quality and maximum efficiency. Various tear sampling methodologies have been described in human studies. In this article, a comprehensive description of an optimized protocol is presented, specifically tailored for extracting tear-related protein information from experimental animal models, especially mice. This method includes the pharmacological stimulation of tear production in 2-month-old mice, followed by sample collection using Schirmer strips and the evaluation of the efficacy and efficiency of the protocol through standard procedures, SDS-PAGE, qPCR, and digital PCR (dPCR). This protocol can be easily adapted for the investigation of the tear protein signature in a variety of experimental paradigms. By establishing an affordable, standardized, and optimized tear sampling protocol for animal models, the aim was to bridge the gap between human and animal research, facilitating translational studies and accelerating advancements in the field of ocular and systemic disease research.
摘要：
泪膜是一种高度动态的生物流体，能够反映病理相关的分子变化，不仅在眼表，而且在其他组织和器官。这种生物流体的分子分析提供了一种非侵入性的方法来诊断或监测疾病，评估药物治疗效果，并确定可能的生物标志物。由于样品体积有限，收集泪液样品需要特定的技能和适当的工具，以确保高质量和最大的效率。在人类研究中已经描述了各种泪液取样方法。在这篇文章中,给出了优化协议的全面描述，专门用于从实验动物模型中提取泪液相关蛋白质信息，尤其是老鼠。该方法包括2个月大的小鼠的泪液产生的药理刺激，然后使用Schirmer试纸条进行样品收集，并通过标准程序评估方案的功效和效率，SDS-PAGE,qPCR,和数字PCR(dPCR)。该方案可以很容易地适用于各种实验范式中的泪液蛋白特征的研究。通过建立一个负担得起的，标准化，和优化的动物模型泪液采样方案，目的是弥合人类和动物研究之间的差距，促进转化研究，加速眼部和全身疾病研究领域的进步。
