美国波士顿大学Sheng Xiao研究团队报道了利用定向照明共聚焦显微镜进行大规模深层组织电压成像技术。相关论文于2024年6月5日发表在《自然-方法学》杂志上。

研究人员开发了一种显微镜，它结合了定向照明和共聚焦选通的独特优势，同时还能最大限度地提高信号检测效率。研究人员通过实验和理论量化了该显微镜在信噪比和串扰减少方面的优势。这种显微镜为实现大尺度和穿透深度的高保真活体电压成像提供了一个多功能解决方案，研究人员在各种成像条件和不同基因编码的电压指示器类别中进行了验证。

据了解，基因编码电压指示器的进步使细胞特异性电压成像成为可能。然而，电压成像所需的千赫兹速率会导致信号减弱。此外，焦外荧光和组织散射产生的背景既影响信噪比，又会引起细胞间的串扰，这使得在高密度标记组织中进行可靠的活体成像极具挑战性。

附：英文原文

Title: Large-scale deep tissue voltage imaging with targeted-illumination confocal microscopy

Author: Xiao, Sheng, Cunningham, William J., Kondabolu, Krishnakanth, Lowet, Eric, Moya, Maria V., Mount, Rebecca A., Ravasio, Cara, Bortz, Emma, Shaw, Dana, Economo, Michael N., Han, Xue, Mertz, Jerome

Issue&Volume: 2024-06-05

Abstract: Voltage imaging with cellular specificity has been made possible by advances in genetically encoded voltage indicators. However, the kilohertz rates required for voltage imaging lead to weak signals. Moreover, out-of-focus fluorescence and tissue scattering produce background that both undermines the signal-to-noise ratio and induces crosstalk between cells, making reliable in vivo imaging in densely labeled tissue highly challenging. We describe a microscope that combines the distinct advantages of targeted illumination and confocal gating while also maximizing signal detection efficiency. The resulting benefits in signal-to-noise ratio and crosstalk reduction are quantified experimentally and theoretically. Our microscope provides a versatile solution for enabling high-fidelity in vivo voltage imaging at large scales and penetration depths, which we demonstrate across a wide range of imaging conditions and different genetically encoded voltage indicator classes.

DOI: 10.1038/s41592-024-02275-w

Source: https://www.nature.com/articles/s41592-024-02275-w