Title | Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators. |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | Chamberland, Simon, Helen H. Yang, Michael M. Pan, Stephen W. Evans, Sihui Guan, Mariya Chavarha, Ying Yang, Charleen Salesse, Haodi Wu, Joseph C. Wu, Thomas R. Clandinin, Katalin Toth, Michael Z. Lin, and François St-Pierre |
Journal | Elife |
Volume | 6 |
Date Published | 2017 07 27 |
ISSN | 2050-084X |
Keywords | Action Potentials, Animals, Drosophila melanogaster, Drosophila Proteins, Female, HEK293 Cells, Humans, Image Processing, Computer-Assisted, Male, Microscopy, Nerve Tissue Proteins, Neurons, Optogenetics, Organ Culture Techniques, Photons, Rats, Sprague-Dawley, Rats, Wistar, Subcellular Fractions, Voltage-Sensitive Dye Imaging |
Abstract | Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in . These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision. |
DOI | 10.7554/eLife.25690 |
Alternate Journal | Elife |
PubMed ID | 28749338 |
PubMed Central ID | PMC5584994 |
Grant List | U01 NS090600 / NS / NINDS NIH HHS / United States P30 NS069375 / NS / NINDS NIH HHS / United States K99 HL133473 / HL / NHLBI NIH HHS / United States MOP-81142 / / CIHR / Canada R21 NS081507 / NS / NINDS NIH HHS / United States S10 RR027431 / RR / NCRR NIH HHS / United States R01 EY022638 / EY / NEI NIH HHS / United States |