小柯机器人

美国加州大学欧文分校Jennifer A. Prescher、Michelle A. Digman等研究人员合作开发出通过相量分析实现的多重生物发光显微镜。相关论文于2022年6月23日在线发表于国际学术期刊《自然—方法学》。

据研究人员介绍，使用荧光素对的生物发光成像是一种成熟的技术，被用于可视化整个组织和整个生物体的生物过程。相比之下，微尺度的应用由于缺乏检测平台和容易解决的探针而受到阻碍。

研究人员通过将生物发光与相量分析相结合来解决了这一限制，相量分析是一种常用于区分光谱相似的荧光团的方法。研究人员建立了一个基于摄像头的显微镜，配备了特殊的光学过滤器，可以直接将相量器的位置分配给独特的荧光素对。六个生物发光报告器在活细胞中很容易被分辨出来，而且读数是定量的和即时的。多重成像也在较长的时间段内进行。生物发光相量仪进一步提供了单细胞中共振能量转移的直接测量，为细胞和分子特征的动态测量创造了条件。生物发光与相量分析的合并填补了成像能力的长期空白，并将促进未来在实时和多个长度尺度上可视化生物事件的努力。

附：英文原文

Title: Multiplexed bioluminescence microscopy via phasor analysis

Author: Yao, Zi, Brennan, Caroline K., Scipioni, Lorenzo, Chen, Hongtao, Ng, Kevin K., Tedeschi, Giulia, Parag-Sharma, Kshitij, Amelio, Antonio L., Gratton, Enrico, Digman, Michelle A., Prescher, Jennifer A.

Issue&Volume: 2022-06-23

Abstract: Bioluminescence imaging with luciferase–luciferin pairs is a well-established technique for visualizing biological processes across tissues and whole organisms. Applications at the microscale, by contrast, have been hindered by a lack of detection platforms and easily resolved probes. We addressed this limitation by combining bioluminescence with phasor analysis, a method commonly used to distinguish spectrally similar fluorophores. We built a camera-based microscope equipped with special optical filters to directly assign phasor locations to unique luciferase–luciferin pairs. Six bioluminescent reporters were easily resolved in live cells, and the readouts were quantitative and instantaneous. Multiplexed imaging was also performed over extended time periods. Bioluminescent phasor further provided direct measures of resonance energy transfer in single cells, setting the stage for dynamic measures of cellular and molecular features. The merger of bioluminescence with phasor analysis fills a long-standing void in imaging capabilities, and will bolster future efforts to visualize biological events in real time and over multiple length scales.

DOI: 10.1038/s41592-022-01529-9

Source: https://www.nature.com/articles/s41592-022-01529-9

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex