BioAFM: Integrated atomic force and light microscopy for mechanochemical cell biology

BioAFM：用于机械化学细胞生物学的集成原子力和光学显微镜

• 批准号: BB/X019934/1
• 负责人: Masanori Mishima
• 金额: $ 97.63万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX019934%2F1
• 关键词: BioAFM; Integrated; atomic; force; light

We explore the environment surrounding us through the combination of multiple senses, i.e., sight, hearing, smell, taste, and touch. Sensing with light is very powerful as it allows us to detect objects without direct contact. However, there are types of information that can't be obtained by sight alone. For example, touch tells us the mechanical properties of an object. By touching someone's hand with your finger, you can feel the texture of the skin and how soft or hard the skin is. You can also find how many fingers exist on that hand and even the bones beneath the skin and flesh. Life science at the scale of molecular machinery, cells and tissues has been heavily relying on light microscopy especially when we try to observe them in their living state. Measurement of the mechanical properties by 'touching' should add another dimension to our sensing of the biological object and provides information that can't be obtained by light microscopy. An atomic force microscope (AFM) is an instrument that achieves such measurements.We integrate an AFM with a light microscope that has two different modes of observation, 1) total internal fluorescence microscopy (TIRFM), which can detect single molecules by fluorescence, and 2) high-resolution 3D fluorescence microscopy (3DFM), which is suitable for observation of thick samples such as cell and tissues. The combination of AFM and TIRFM will allow us to visualize the live dynamics of the assembly of biomolecular machines that play key roles in the transport/traffic inside the cell. AFM+3DFM enables us to measure various mechanical properties of a single cell of interest in a complex multicellular environment by using a fluorescent marker as a guide. Our proposed and future BioAFM experiments will give new insights into cellular and molecular mechanisms of cell and tissue morphogenesis and other biological processes, in which mechanical force plays a key role.

我们通过视觉、听觉、嗅觉、味觉和触觉等多种感官的结合来探索周围的环境。光感应是非常强大的，因为它可以让我们在没有直接接触的情况下探测到物体。然而，有些类型的信息是不能仅凭视觉获得的。例如，触摸告诉我们物体的机械特性。通过用手指触摸别人的手，你可以感觉到皮肤的质地和皮肤的软硬程度。你还可以找到那只手上有多少根手指，甚至是皮肤和肉下面的骨头。分子机械、细胞和组织的生命科学在很大程度上依赖于光学显微镜，特别是当我们试图观察它们的生活状态时。通过“触摸”来测量机械性能应该为我们对生物物体的感知增加另一个维度，并提供光显微镜无法获得的信息。原子力显微镜（AFM）就是实现这种测量的仪器。我们将AFM与光学显微镜相结合，光学显微镜具有两种不同的观察模式，1)全内荧光显微镜（TIRFM），可以通过荧光检测单个分子，2)高分辨率三维荧光显微镜（3DFM），适用于观察细胞和组织等厚样品。AFM和TIRFM的结合将使我们能够可视化生物分子机器组装的动态，这些机器在细胞内的运输/交通中起着关键作用。AFM+3DFM使我们能够在复杂的多细胞环境中通过使用荧光标记作为指导来测量单个感兴趣细胞的各种机械特性。我们提出的和未来的生物afm实验将为细胞和组织形态发生以及其他生物过程的细胞和分子机制提供新的见解，其中机械力起着关键作用。