小柯机器人

美国斯坦福大学Michael Z. Lin研究团队开发出用于脑部扩展电记录的正调电压指示器。2023年7月6日，《自然—方法学》杂志在线发表了这一最新研究成果。

研究人员报告了ASAP系列基因编码的电压指示器（GEVI）的工程化，其通过反转荧光-电压关系来提高光稳定性。由此产生的两个GEVI，ASAP4b和ASAP4e，对100-mV的去极化反应，荧光增加≥180%，而亲本ASAP3的荧光减少50%。使用标准的显微镜设备，ASAP4e能在几分钟内对小鼠的脉冲进行单次检测。与以前用于单光子电压记录的GEVI不同，ASAP4b和ASAP4e在双光子照明下也表现良好。通过对电压和钙的同时成像，研究人员表明ASAP4b和ASAP4e可以识别位置细胞，并以比常用的钙指示器更好的时间分辨率检测电压脉冲。因此，ASAP4b和ASAP4e将电压成像的能力扩展到标准的单光子和双光子显微镜，同时改善了电压记录的持续时间。

据介绍，GEVI能够对大脑中的电信号进行光学记录，提供钙指示器不可能提供的亚阈值灵敏度和时间分辨率。然而，用同一个GEVI进行长时间的单光子和双光子电压成像还没有被证明。

附：英文原文

Title: A positively tuned voltage indicator for extended electrical recordings in the brain

Author: Evans, S. Wenceslao, Shi, Dong-Qing, Chavarha, Mariya, Plitt, Mark H., Taxidis, Jiannis, Madruga, Blake, Fan, Jiang Lan, Hwang, Fuu-Jiun, van Keulen, Siri C., Suomivuori, Carl-Mikael, Pang, Michelle M., Su, Sharon, Lee, Sungmoo, Hao, Yukun A., Zhang, Guofeng, Jiang, Dongyun, Pradhan, Lagnajeet, Roth, Richard H., Liu, Yu, Dorian, Conor C., Reese, Austin L., Negrean, Adrian, Losonczy, Attila, Makinson, Christopher D., Wang, Sui, Clandinin, Thomas R., Dror, Ron O., Ding, Jun B., Ji, Na, Golshani, Peyman, Giocomo, Lisa M., Bi, Guo-Qiang, Lin, Michael Z.

Issue&Volume: 2023-07-06

Abstract: Genetically encoded voltage indicators (GEVIs) enable optical recording of electrical signals in the brain, providing subthreshold sensitivity and temporal resolution not possible with calcium indicators. However, one- and two-photon voltage imaging over prolonged periods with the same GEVI has not yet been demonstrated. Here, we report engineering of ASAP family GEVIs to enhance photostability by inversion of the fluorescence–voltage relationship. Two of the resulting GEVIs, ASAP4b and ASAP4e, respond to 100-mV depolarizations with ≥180% fluorescence increases, compared with the 50% fluorescence decrease of the parental ASAP3. With standard microscopy equipment, ASAP4e enables single-trial detection of spikes in mice over the course of minutes. Unlike GEVIs previously used for one-photon voltage recordings, ASAP4b and ASAP4e also perform well under two-photon illumination. By imaging voltage and calcium simultaneously, we show that ASAP4b and ASAP4e can identify place cells and detect voltage spikes with better temporal resolution than commonly used calcium indicators. Thus, ASAP4b and ASAP4e extend the capabilities of voltage imaging to standard one- and two-photon microscopes while improving the duration of voltage recordings.

DOI: 10.1038/s41592-023-01913-z

Source: https://www.nature.com/articles/s41592-023-01913-z

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