Several techniques have been developed over the past few decades to assess the mechanical properties of biological samples, which has fueled a rapid growth in the fields of biophysics, bioengineering, and mechanobiology. In this context, Brillouin optical spectroscopy has long been known as an intriguing modality for noncontact material characterization. However, limited by speed and sample damage, it had not translated into a viable imaging modality for biomedically relevant materials. Recently, based on a novel spectroscopy strategy that substantially improves the speed of Brillouin measurement, confocal Brillouin microscopy has emerged as a unique complementary tool to traditional methods as it allows noncontact, nonperturbative, label-free measurements of material mechanical properties. The feasibility and potential of this innovative technique at both the cell and tissue level have been extensively demonstrated over the past decade. As Brillouin technology is rapidly recognized, a standard approach for building and operating Brillouin microscopes is required to facilitate the widespread adoption of this technology. In this protocol, we aim to establish a robust approach for instrumentation, and data acquisition and analysis. By carefully following this protocol, we expect that a Brillouin instrument can be built in 5–9 days by a person with basic optics knowledge and alignment experience; the data acquisition as well as postprocessing can be accomplished within 2–8 h.

在过去的几十年里，已经开发出多种技术来评估生物样品的机械特性，这推动了生物物理学、生物工程和力学生物学领域的快速发展。在这种情况下，布里渊光谱长期以来一直被认为是一种用于非接触式材料表征的有趣模式。然而，受速度和样品损坏的限制，它还没有转化为生物医学相关材料的可行成像方式。最近，基于大幅提高布里渊测量速度的新型光谱策略，共聚焦布里渊显微镜已成为传统方法的独特补充工具，因为它允许对材料机械性能进行非接触、非微扰、无标记测量。在过去十年中，这种创新技术在细胞和组织水平上的可行性和潜力已得到广泛证明。随着布里渊技术迅速得到认可，需要一种构建和操作布里渊显微镜的标准方法来促进该技术的广泛采用。在本协议中，我们的目标是为仪器、数据采集和分析建立一个强大的方法。通过仔细遵循此协议，我们预计布里渊仪器可以由具有基本光学知识和对准经验的人员在 5-9 天内构建；数据采集​​和后处理可以在 2-8 小时内完成。需要一种用于构建和操作布里渊显微镜的标准方法，以促进该技术的广泛采用。在本协议中，我们的目标是为仪器、数据采集和分析建立一个强大的方法。通过仔细遵循此协议，我们预计布里渊仪器可以由具有基本光学知识和对准经验的人员在 5-9 天内构建；数据采集​​和后处理可以在 2-8 小时内完成。需要一种用于构建和操作布里渊显微镜的标准方法，以促进该技术的广泛采用。在本协议中，我们的目标是为仪器、数据采集和分析建立一个强大的方法。通过仔细遵循此协议，我们预计布里渊仪器可以由具有基本光学知识和对准经验的人员在 5-9 天内构建；数据采集​​和后处理可以在 2-8 小时内完成。我们期望具有基本光学知识和对准经验的人可以在 5-9 天内构建布里渊仪器；数据采集​​和后处理可以在 2-8 小时内完成。我们期望具有基本光学知识和对准经验的人可以在 5-9 天内构建布里渊仪器；数据采集​​和后处理可以在 2-8 小时内完成。