High-throughput synapse-resolving two-photon fluorescence microendoscopy for deep-brain volumetric imaging in vivo
- University of California, Berkeley, United States
- Janelia Research Campus, Howard Hughes Medical Institute, United States
- National Institute on Drug Abuse, United States
- Johns Hopkins University School of Medicine, United States
Abstract
Optical imaging has become a powerful tool for studying brains in vivo. The opacity of adult brains makes microendoscopy, with an optical probe such as a gradient index (GRIN) lens embedded into brain tissue to provide optical relay, the method of choice for imaging neurons and neural activity in deeply buried brain structures. Incorporating a Bessel focus scanning module into two-photon fluorescence microendoscopy, we extended the excitation focus axially and improved its lateral resolution. Scanning the Bessel focus in 2D, we imaged volumes of neurons at high-throughput while resolving fine structures such as synaptic terminals. We applied this approach to the volumetric anatomical imaging of dendritic spines and axonal boutons in the mouse hippocampus, and functional imaging of GABAergic neurons in the mouse lateral hypothalamus in vivo.
Data availability
Almost all data needed to evaluate the conclusions in the paper are present in the paper or the supplementary materials; Raw image data for Figs. 2, 4 & 9 are available from Dryad, 10.5061/dryad.pr4t978
Article and author information
Author details
Joshua Tate Dudman
Funding
Howard Hughes Medical Institute
- Guanghan Meng
- Yajie Liang
- Wan-chen Jiang
- Rongwen Lu
- Joshua Tate Dudman
- Na Ji
National Institute of Neurological Disorders and Stroke
- Guanghan Meng
- Na Ji
National Institute on Drug Abuse
- Sarah Sarsfield
- Yeka Aponte
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All animal experiments were conducted according to the United States National Institutes of Health guidelines for animal research. Procedures and protocols were approved by the Institutional Animal Care and Use Committee at Janelia Research Campus, Howard Hughes Medical Institute (protocol number: 16-147)
Copyright
© 2019, Meng et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 11,511
- views
-
- 1,617
- downloads
-
- 78
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
High-throughput synapse-resolving two-photon fluorescence microendoscopy for deep-brain volumetric imaging in vivo
eLife 8:e40805.
https://doi.org/10.7554/eLife.40805
Further reading
-
- Computational and Systems Biology
- Neuroscience
We determined the intersubject association between the rhythmic entrainment abilities of human subjects during a synchronization-continuation tapping task (SCT) and the macro- and microstructural properties of their superficial (SWM) and deep (DWM) white matter. Diffusion-weighted images were obtained from 32 subjects who performed the SCT with auditory or visual metronomes and five tempos ranging from 550 to 950 ms. We developed a method to determine the density of short-range fibers that run underneath the cortical mantle, interconnecting nearby cortical regions (U-fibers). Notably, individual differences in the density of U-fibers in the right audiomotor system were correlated with the degree of phase accuracy between the stimuli and taps across subjects. These correlations were specific to the synchronization epoch with auditory metronomes and tempos around 1.5 Hz. In addition, a significant association was found between phase accuracy and the density and bundle diameter of the corpus callosum, forming an interval-selective map where short and long intervals were behaviorally correlated with the anterior and posterior portions of the corpus callosum. These findings suggest that the structural properties of the SWM and DWM in the audiomotor system support the tapping synchronization abilities of subjects, as cortical U-fiber density is linked to the preferred tapping tempo and the bundle properties of the corpus callosum define an interval-selective topography.
-
- Neuroscience
Real-world actions often comprise a series of movements that cannot be entirely planned before initiation. When these actions are executed rapidly, the planning of multiple future movements needs to occur simultaneously with the ongoing action. How the brain solves this task remains unknown. Here, we address this question with a new sequential arm reaching paradigm that manipulates how many future reaches are available for planning while controlling execution of the ongoing reach. We show that participants plan at least two future reaches simultaneously with an ongoing reach. Further, the planning processes of the two future reaches are not independent of one another. Evidence that the planning processes interact is twofold. First, correcting for a visual perturbation of the ongoing reach target is slower when more future reaches are planned. Second, the curvature of the current reach is modified based on the next reach only when their planning processes temporally overlap. These interactions between future planning processes may enable smooth production of sequential actions by linking individual segments of a long sequence at the level of motor planning.
