在活细胞中塑造光的高空间分辨率光遗传学激活的倏逝焦点
Shaping an evanescent focus of light for high spatial resolution optogenetic activations in live cells
AI总结：
提出利用DMD在高数值孔径物镜中创建并移动聚焦倏逝斑的方法，实现亚微米横向尺寸和约100 nm轴向深度的三维光限制，显著提升了活细胞CRY2-CIBN光遗传学系统的空间激活分辨率。
摘要：
在三维空间中限制光照明对各种应用而言是一项挑战。其中，为细胞生物学活体实验开发的光遗传学方法将受益于这种局部照明，因为它能提高扩散型光敏蛋白的空间分辨率，从而在特定亚细胞器中产生空间受限的生物反应。在此，我们描述了一种利用数字微镜器件（DMD），在高数值孔径显微镜物镜视场内创建并移动聚焦倏逝斑的方法。我们证明，在校正光学像差后，光被限制在横向亚微米尺寸和约100 nm轴向深度的光斑内，使得照明体积远小于标准传播焦点产生的体积。在光遗传学系统 CRY2-CIBN 上，该倏逝焦点足以引发比传播焦点更强烈且更局部的招募，从而提高了其激活图谱的分辨率。
英文摘要：
Confining light illumination in the three dimensions of space is a challenge for various applications. Among these, optogenetic methods developed for live experiments in cell biology would benefit from such a localized illumination as it would improve the spatial resolution of diffusive photosensitive proteins leading to spatially constrained biological responses in specific subcellular organelles. Here, we describe a method to create and move a focused evanescent spot across the field of view of a high numerical aperture microscope objective, using a digital micro-mirror device (DMD). We show that, after correcting the optical aberrations, light is confined within a spot of sub-micron lateral size and $\sim$100~nm axial depth, resulting in a volume of illumination drastically smaller than the one generated by a standard propagative focus. This evanescent focus is sufficient to induce a more intense and localized recruitment compared to a propagative focus on the optogenetic system CRY2-CIBN, improving the resolution of its pattern of activation.
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