Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published September 28, 2021 | Published + Supplemental Material
Journal Article Open

Light-guided sectioning for precise in situ localization and tissue interface analysis for brain-implanted optical fibers and GRIN lenses


Optical implants to control and monitor neuronal activity in vivo have become foundational tools of neuroscience. Standard two-dimensional histology of the implant location, however, often suffers from distortion and loss during tissue processing. To address that, we developed a three-dimensional post hoc histology method called "light-guided sectioning" (LiGS), which preserves the tissue with its optical implant in place and allows staining and clearing of a volume up to 500 μm in depth. We demonstrate the use of LiGS to determine the precise location of an optical fiber relative to a deep brain target and to investigate the implant-tissue interface. We show accurate cell registration of ex vivo histology with single-cell, two-photon calcium imaging, obtained through gradient refractive index (GRIN) lenses, and identify subpopulations based on immunohistochemistry. LiGS provides spatial information in experimental paradigms that use optical fibers and GRIN lenses and could help increase reproducibility through identification of fiber-to-target localization and molecular profiling.

Additional Information

© 2021 The Author(s). This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Received 25 November 2020, Revised 22 June 2021, Accepted 31 August 2021, Available online 29 September 2021. Published: September 28, 2021. We would like to thank the Gradinaru laboratory and Prof. Justin Bois for helpful discussions, the Caltech Neurotechnology Laboratory for the two-photon microscope support, the Caltech Biological Imaging Center for the training and use of the Airyscan confocal microscope, and Dr. Ariane Helou for editorial comments. Professor Gradinaru is a Heritage Principal Investigator supported by the Heritage Medical Research Institute; this work was also funded by the NIH Director's New Innovator IDP20D017782-01, the NSF NeuroNex Technology Hub 1707316, Presidential Early Career Awards for Scientists and Engineers (V.G.), NIH Brain Research through Advancing Innovative Neurotechnologies RF1MH117069, and the Beckman Institute's Resource Center on CLARITY, Optogenetics, and Vector Engineering (CLOVER). We would also like to thank CLOVER for technology development and broad dissemination (https://beckmaninstitute.caltech.edu/clover.shtml). A.K. is supported by a Caltech Biology and Biological Engineering department postdoctoral fellowship and the Hebrew University Postdoctoral Fellowship for Women, Israel. A.G. is a Good Ventures Fellow of the Life Sciences Research Foundation. P.K. is supported by the Swiss National Science Foundation (P2EZP3-181896). Author contributions. Conceptualization, A.K., A.G., and V.G.; methodology, A.K. and A.G.; software, A.K., A.G., J.R.C., P.K., and D.A.W; investigation, A.K.; resources, A.K., A.G., M.J.J., J.E.R, J.R.C., X.C., D.A.W., and V.G.; writing – original draft, A.K., and A.G.; writing – review & editing, A.K., A.G., M.J.J., J.E.R, J.R.C., X.C., P.K., D.A.W, and V.G.; visualization, A.K. and A.G.; supervision, V.G. The authors declare no competing interests. Data and code availability The datasets supporting the current study have not been deposited in a public repository but are available from the corresponding author upon reasonable request. All original code has been deposited at github.com and is publicly available as of the date of publication. DOIs are listed in the key resources table. Any additional information required to reanalyze the data reported in this paper is available from the lead contact upon request.

Attached Files

Published - 1-s2.0-S2211124721011979-main.pdf

Supplemental Material - 1-s2.0-S2211124721011979-mmc1.pdf

Supplemental Material - 1-s2.0-S2211124721011979-mmc2.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc3.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc4.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc5.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc6.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc7.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc8.mp4

Supplemental Material - 1-s2.0-S2211124721011979-mmc9.mp4


Files (180.2 MB)
Name Size Download all
6.2 MB Preview Download
11.3 MB Download
2.2 MB Preview Download
15.7 MB Download
37.0 MB Download
13.9 MB Download
13.0 MB Download
4.1 MB Download
35.3 MB Download
41.5 MB Download

Additional details

August 22, 2023
December 22, 2023