High-throughput synapse-resolving two-photon fluorescence microendoscopy for deep-brain volumetric imaging in vivo

  1. Guanghan Meng
  2. Yajie Liang
  3. Sarah Sarsfield
  4. Wan-chen Jiang
  5. Rongwen Lu
  6. Joshua Tate Dudman
  7. Yeka Aponte
  8. Na Ji  Is a corresponding author
  1. University of California, Berkeley, United States
  2. Janelia Research Campus, Howard Hughes Medical Institute, United States
  3. National Institute on Drug Abuse, United States
  4. 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

The following data sets were generated

Article and author information

Author details

  1. Guanghan Meng

    Department of Physics, University of California, Berkeley, Berkeley, United States
    Competing interests
    No competing interests declared.
  2. Yajie Liang

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    No competing interests declared.
  3. Sarah Sarsfield

    Neuronal Circuits and Behavior Unit, National Institute on Drug Abuse, Baltimore, United States
    Competing interests
    No competing interests declared.
  4. Wan-chen Jiang

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    No competing interests declared.
  5. Rongwen Lu

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    Rongwen Lu, The Bessel focus scanning intellectual property has been licensed to Thorlabs, Inc. by HHMI.
  6. Joshua Tate Dudman

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4436-1057
  7. Yeka Aponte

    Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5967-2579
  8. Na Ji

    Department of Physics, University of California, Berkeley, Berkeley, United States
    For correspondence
    jina@berkeley.edu
    Competing interests
    Na Ji, The Bessel focus scanning intellectual property has been licensed to Thorlabs, Inc. by HHMI.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5527-1663

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

  • 12,026
    views
  • 1,662
    downloads
  • 84
    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)

  1. Guanghan Meng
  2. Yajie Liang
  3. Sarah Sarsfield
  4. Wan-chen Jiang
  5. Rongwen Lu
  6. Joshua Tate Dudman
  7. Yeka Aponte
  8. Na Ji
(2019)
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

Share this article

https://doi.org/10.7554/eLife.40805

Further reading

    1. Neuroscience
    Sven Ohl, Martin Rolfs
    Research Article

    Detecting causal relations structures our perception of events in the world. Here, we determined for visual interactions whether generalized (i.e. feature-invariant) or specialized (i.e. feature-selective) visual routines underlie the perception of causality. To this end, we applied a visual adaptation protocol to assess the adaptability of specific features in classical launching events of simple geometric shapes. We asked observers to report whether they observed a launch or a pass in ambiguous test events (i.e. the overlap between two discs varied from trial to trial). After prolonged exposure to causal launch events (the adaptor) defined by a particular set of features (i.e. a particular motion direction, motion speed, or feature conjunction), observers were less likely to see causal launches in subsequent ambiguous test events than before adaptation. Crucially, adaptation was contingent on the causal impression in launches as demonstrated by a lack of adaptation in non-causal control events. We assessed whether this negative aftereffect transfers to test events with a new set of feature values that were not presented during adaptation. Processing in specialized (as opposed to generalized) visual routines predicts that the transfer of visual adaptation depends on the feature similarity of the adaptor and the test event. We show that the negative aftereffects do not transfer to unadapted launch directions but do transfer to launch events of different speeds. Finally, we used colored discs to assign distinct feature-based identities to the launching and the launched stimulus. We found that the adaptation transferred across colors if the test event had the same motion direction as the adaptor. In summary, visual adaptation allowed us to carve out a visual feature space underlying the perception of causality and revealed specialized visual routines that are tuned to a launch’s motion direction.

    1. Neuroscience
    Gergely F Turi, Sasa Teng ... Yueqing Peng
    Research Article

    Synchronous neuronal activity is organized into neuronal oscillations with various frequency and time domains across different brain areas and brain states. For example, hippocampal theta, gamma, and sharp wave oscillations are critical for memory formation and communication between hippocampal subareas and the cortex. In this study, we investigated the neuronal activity of the dentate gyrus (DG) with optical imaging tools during sleep-wake cycles in mice. We found that the activity of major glutamatergic cell populations in the DG is organized into infraslow oscillations (0.01–0.03 Hz) during NREM sleep. Although the DG is considered a sparsely active network during wakefulness, we found that 50% of granule cells and about 25% of mossy cells exhibit increased activity during NREM sleep, compared to that during wakefulness. Further experiments revealed that the infraslow oscillation in the DG was correlated with rhythmic serotonin release during sleep, which oscillates at the same frequency but in an opposite phase. Genetic manipulation of 5-HT receptors revealed that this neuromodulatory regulation is mediated by Htr1a receptors and the knockdown of these receptors leads to memory impairment. Together, our results provide novel mechanistic insights into how the 5-HT system can influence hippocampal activity patterns during sleep.