■无标签显微镜的进步可以提供实时,非侵入性成像,具有独特的造影剂来源和自动化标准化分析,以表征异质和动态的生物过程。这些工具将克服广泛使用的破坏性方法的挑战(例如,组织学,流式细胞术)或缺乏细胞分辨率(例如,基于平板的测定,整个动物生物发光成像)。
■此观点旨在(1)证明无标签显微镜在不受干扰的系统中随时间和空间跟踪异质细胞功能的必要性,以及(2)建议有关仪器的改进,图像分析,和图像解释来满足这些需求。
■三个关键研究领域(癌症研究,自身免疫性疾病,以及组织和细胞工程)被认为支持对无标记显微镜的需求,以表征生物系统内的异质性和动力学。基于优势(例如,多种分子对比来源,非侵入性监测)和弱点(例如,成像深度,图像解释)几种无标签显微镜模式,建议对未来的成像系统进行改进。
■仪器的改进,包括提高分辨率和成像速度的策略,图像分析工具的标准化和集中化,和强大的数据验证和解释将扩大无标签显微镜的应用,以研究异质和动态的生物系统。
UNASSIGNED: Advancements in label-free microscopy could provide real-time, non-invasive imaging with unique sources of contrast and automated standardized analysis to characterize heterogeneous and dynamic biological processes. These tools would overcome challenges with widely used methods that are destructive (e.g., histology, flow cytometry) or lack cellular resolution (e.g., plate-based assays, whole animal bioluminescence imaging).
UNASSIGNED: This perspective aims to (1) justify the need for label-free microscopy to track heterogeneous cellular functions over time and space within unperturbed systems and (2) recommend improvements regarding instrumentation, image analysis, and image interpretation to address these needs.
UNASSIGNED: Three key research areas (cancer research, autoimmune disease, and tissue and cell engineering) are considered to support the need for label-free microscopy to characterize heterogeneity and dynamics within biological systems. Based on the strengths (e.g., multiple sources of molecular contrast, non-invasive monitoring) and weaknesses (e.g., imaging depth, image interpretation) of several label-free microscopy modalities, improvements for future imaging systems are recommended.
UNASSIGNED: Improvements in instrumentation including strategies that increase resolution and imaging speed, standardization and centralization of image analysis tools, and robust data validation and interpretation will expand the applications of label-free microscopy to study heterogeneous and dynamic biological systems.