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

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

doi:10.1038/s41592-023-01913-z

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

Nat Methods. 2023 Jul;20(7):1104-1113. doi: 10.1038/s41592-023-01913-z. Epub 2023 Jul 6.

Authors

S Wenceslao Evans^#¹, Dong-Qing Shi^#², Mariya Chavarha^#³, Mark H Plitt¹, Jiannis Taxidis^{4

5

6

5

6}, Blake Madruga⁴, Jiang Lan Fan⁷, Fuu-Jiun Hwang⁸, Siri C van Keulen⁹, Carl-Mikael Suomivuori⁹, Michelle M Pang¹, Sharon Su¹, Sungmoo Lee¹, Yukun A Hao^{1

3}, Guofeng Zhang¹, Dongyun Jiang¹, Lagnajeet Pradhan¹, Richard H Roth⁸, Yu Liu^{8

10}, Conor C Dorian⁴, Austin L Reese¹¹, Adrian Negrean¹², Attila Losonczy^{12

13

14}, Christopher D Makinson^{11

15}, Sui Wang¹⁰, Thomas R Clandinin¹, Ron O Dror^{9

16

17

18}, Jun B Ding^{8

19}, Na Ji^{5

20

21

22

23}, Peyman Golshani^{4

24}, Lisa M Giocomo¹, Guo-Qiang Bi^{2

25}, Michael Z Lin^{26

27

28}

Affiliations

¹ Department of Neurobiology, Stanford University Medical Center, Stanford, CA, USA.
² Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.
³ Department of Bioengineering, Stanford University, Stanford, CA, USA.
⁴ Department of Neurology, UCLA David Geffen School of Medicine, Los Angeles, CA, USA.
⁵ Program in Neurosciences & Mental Health, Hospital for Sick Children, Toronto, Ontario, Canada.
⁶ Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
⁷ UC Berkeley/UCSF Joint Program in Bioengineering, University of California Berkeley, Berkeley, CA, USA.
⁸ Department of Neurosurgery, Stanford University Medical Center, Stanford, CA, USA.
⁹ Department of Computer Science, Stanford University, Stanford, CA, USA.
¹⁰ Department of Ophthalmology, Stanford University Medical Center, Stanford, CA, USA.
¹¹ Institute for Genomic Medicine, Columbia University, New York, NY, USA.
¹² Department of Neuroscience, Columbia University, New York, NY, USA.
¹³ Mortimer B. Zuckerman Mind Brain Behavior Institute, New York, NY, USA.
¹⁴ Kavli Institute for Brain Science, New York, NY, USA.
¹⁵ Department of Neurology, Columbia University, New York, NY, USA.
¹⁶ Department of Structural Biology, Stanford University School of Medicine, Stanford, CA, USA.
¹⁷ Institute for Computational and Mathematical Engineering, Stanford University, Stanford, CA, USA.
¹⁸ Department of Molecular and Cellular Physiology, Stanford University, Stanford, USA.
¹⁹ Department of Neurology and Neurological Sciences, Stanford University Medical Center, Stanford, CA, USA.
²⁰ Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, USA.
²¹ Department of Physics, University of California Berkeley, Berkeley, CA, USA.
²² Helen Wills Neuroscience Institute, University of California Berkeley, Berkeley, CA, USA.
²³ Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
²⁴ Semel Institute for Neuroscience and Human Behavior, UCLA David Geffen School of Medicine, Los Angeles, CA, USA.
²⁵ Interdisciplinary Center for Brain Information, Brain Cognition and Brain Disease Institute, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
²⁶ Department of Neurobiology, Stanford University Medical Center, Stanford, CA, USA. mzlin@stanford.edu.
²⁷ Department of Bioengineering, Stanford University, Stanford, CA, USA. mzlin@stanford.edu.
²⁸ Department of Chemical and Systems Biology, Stanford University, Stanford, USA. mzlin@stanford.edu.

^# Contributed equally.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Brain*
Calcium*
Lighting
Mice
Microscopy
Photons

Substances

Calcium

Abstract

Publication types

MeSH terms

Substances

Grants and funding