A silent two-photon imaging system for studying in vivo auditory neuronal functions

Xindong Song; Yueqi Guo; Chenggang Chen; Xiaoqin Wang

doi:10.1038/s41377-022-00783-y

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A silent two-photon imaging system for studying in vivo auditory neuronal functions

Articles

- A silent two-photon imaging system for studying in vivo auditory neuronal functions
- Light: Science & Applications Vol. 11, Issue 5, Pages: 888-900(2022)
- Affiliations：
  
  Laboratory of Auditory Neurophysiology, Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
- Author bio：
  
  Xindong Song (songxindong@jhmi.edu)
  Xiaoqin Wang (xiaoqin.wang@jhu.edu)
- Funds：
- DOI：10.1038/s41377-022-00783-y
  CLC：
- Published：31 May 2022，
  
  Published Online：14 April 2022，
  
  Received：25 October 2021，
  
  Revised：28 March 2022，
  
  Accepted：30 March 2022
- Accepted：
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Song, X. D. et al. A silent two-photon imaging system for studying in vivo auditory neuronal functions. Light: Science & Applications, 11, 888-900 (2022).
DOI：

Song, X. D. et al. A silent two-photon imaging system for studying in vivo auditory neuronal functions. Light: Science & Applications, 11, 888-900 (2022). DOI： 10.1038/s41377-022-00783-y.

摘要

Abstract

Two-photon laser-scanning microscopy has become an essential tool for imaging neuronal functions in vivo and has been applied to different parts of the neural system

including the auditory system. However

many components of a two-photon microscope

such as galvanometer-based laser scanners

generate mechanical vibrations and thus acoustic artifacts

making it difficult to interpret auditory responses from recorded neurons. Here

we report the development of a silent two-photon imaging system and its applications in the common marmoset (

Callithrix Jacchus

)

a non-human primate species sharing a similar hearing range with humans. By utilizing an orthogonal pair of acousto-optical deflectors (AODs)

full-frame raster scanning at video rate was achieved without introducing mechanical vibrations. Imaging depth can be optically controlled by adjusting the chirping speed on the AODs without any mechanical motion along the Z-axis. Furthermore

all other sound-generating components of the system were acoustically isolated

leaving the noise floor of the working system below the marmoset's hearing threshold. Imaging with the system in awake marmosets revealed many auditory cortex neurons that exhibited maximal responses at low sound levels

which were not possible to study using traditional two-photon imaging systems. This is the first demonstration of a silent two-photon imaging system that is capable of imaging auditory neuronal functi

ons in vivo without acoustic artifacts. This capacity opens new opportunities for a better understanding of auditory functions in the brain and helps isolate animal behavior from microscope-generated acoustic interference.

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references

Ohki, K. et al. Functional imaging with cellular resolution reveals precise micro-architecture in visual cortex.Nature433, 597–603 (2005)..

Sato, T. R. et al. The functional microarchitecture of the mouse barrel cortex.PLoS Biol.5, e189 (2007)..

Dombeck, D. A., Graziano, M. S.&Tank, D. W. Functional clustering of neurons in motor cortex determined by cellular resolution imaging in awake behaving mice.J. Neurosci.29, 13751–13760 (2009)..

Deneux, T. et al. Context-dependent signaling of coincident auditory and visual events in primary visual cortex.eLife8, e44006 (2019)..

Ceballo, S. et al. Targeted cortical manipulation of auditory perception.Neuron104, 1168–1179. e5 (2019)..

Issa, J. B. et al. Multiscale optical Ca2+imaging of tonal organization in mouse auditory cortex.Neuron83, 944–959 (2014)..

Barnstedt, O. et al. Functional microarchitecture of the mouse dorsal inferior colliculus revealed through in vivo two-photon calcium imaging.J. Neurosci.35, 10927–10939 (2015)..

Panniello, M. et al. Local and global spatial organization of interaural level difference and frequency preferences in auditory cortex.Cereb. Cortex28, 350–369 (2018)..

Tischbirek, C. H. et al. In vivo functional mapping of a cortical column at single-neuron resolution.Cell Rep.27, 1319–1326. e5 (2019)..

Xin, Y. et al. Sensory-to-category transformation via dynamic reorganization of ensemble structures in mouse auditory cortex.Neuron103, 909–921. e6 (2019)..

Romero, S. et al. Cellular and widefield imaging of sound frequency organization in primary and higher order fields of the mouse auditory cortex.Cereb. Cortex30, 1603–1622 (2020)..

Gaucher, Q. et al. Complexity of frequency receptive fields predicts tonotopic variability across species.eLife9, e53462 (2020)..

Bullen, A., Patel, S. S.&Saggau, P. High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.Biophysical J.73, 477–491 (1997)..

Lechleiter, J. D., Lin, D. T.&Sieneart, I. Multi-photon laser scanning microscopy using an acoustic optical deflector.Biophysical J.83, 2292–2299 (2002)..

Nadella, K. M. N. S. et al. Random-access scanning microscopy for 3D imaging in awake behaving animals.Nat. Methods13, 1001–1004 (2016)..

Szalay, G. et al. Fast 3D imaging of spine, dendritic, and neuronal assemblies in behaving animals.Neuron92, 723–738 (2016)..

Osmanski, M. S.&Wang, X. Q. Measurement of absolute auditory thresholds in the common marmoset (Callithrix jacchus).Hearing Res.277, 127–133 (2011)..

Song, X. D. et al. Complex pitch perception mechanisms are shared by humans and a New World monkey.Proc. Natl. Acad. Sci. USA113, 781–786 (2016)..

Miller, C. T. et al. Marmosets: a neuroscientific model of human social behavior.Neuron90, 219–233 (2016)..

Song, X. D. et al. Functional maps of the primate cortex revealed by through-skull wide-field optical imaging. Preprint athttps://doi.org/10.1101/2020.12.05.413047https://doi.org/10.1101/2020.12.05.413047(2020).

Tani, T. et al. Sound frequency representation in the auditory cortex of the common marmoset visualized using optical intrinsic signal imaging.eNeuro5, ENEURO. 0078-18.2018 (2018)..

Zeng, H. H. et al. Local homogeneity of tonotopic organization in the primary auditory cortex of marmosets.Proc. Natl Acad. Sci. USA116, 3239–3244 (2019)..

Moore, B. C. J.&Glasberg, B. R. A revision of Zwicker's loudness model.Acust. U. Acta Acust.82, 335–345 (1996)..

Osmanski, M. S. et al. Frequency discrimination in the common marmoset (Callithrix jacchus).Hearing Res.341, 1–8 (2016)..

Radziwon, K. E. et al. Behaviorally measured audiograms and gap detection thresholds in CBA/CaJ mice.J. Comp. Physiol. A195, 961–969 (2009)..

Kelly, J. B., Kavanagh, G. L.&Dalton, J. C. H. Hearing in the ferret (Mustela putorius): thresholds for pure tone detection.Hearing Res.24, 269–275 (1986)..

Borg, E. Auditory thresholds in rats of different age and strain. A behavioral and electrophysiological study.Hearing Res.8, 101–115 (1982)..

Zwicker, E., Flottorp, G.&Stevens, S. S. Critical band width in loudness summation.J. Acoustical Soc. Am.29, 548–557 (1957)..

Osmanski, M. S., Song, X. D.&Wang, X. Q. The role of harmonic resolvability in pitch perception in a vocal nonhuman primate, the common marmoset (Callithrix jacchus).J. Neurosci.33, 9161–9168 (2013)..

Bandyopadhyay, S., Shamma, S. A.&Kanold, P. O. Dichotomy of functional organization in the mouse auditory cortex.Nat. Neurosci.13, 361–368 (2010)..

Zheng, Q. Y., Johnson, K. R.&Erway, L. C. Assessment of hearing in 80 inbred strains of mice by ABR threshold analyses.Hearing Res.130, 94–107 (1999)..

Bowen, Z., Winkowski, D. E.&Kanold, P. O. Functional organization of mouse primary auditory cortex in adult C57BL/6 and F1 (CBAxC57) mice.Sci. Rep.10, 10905 (2020)..

Mikaelian, D. O., Warfield, D.&Norris, O. Genetic progressive hearing loss in the C57/M6 mouse:relation of behaviorial responses to cochlear anatomy.Acta Oto-laryngologica77, 327–334 (1974)..

Grewe, B. F. et al. High-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision.Nat. Methods7, 399–405 (2010)..

Pnevmatikakis, E. A.&Giovannucci, A. NoRMCorre: an online algorithm for piecewise rigid motion correction of calcium imaging data.J. Neurosci. Methods291, 83–94 (2017)..

Vučinić, D.&Sejnowski, T. J. A compact multiphoton 3D imaging system for recording fast neuronal activity.PLoS One2, e699 (2007)..

Chen, X. W. et al. Functional mapping of single spines in cortical neurons in vivo.Nature475, 501–505 (2011)..

Zheng, T. et al. Visualization of brain circuits using two-photon fluorescence micro-optical sectioning tomography.Opt. Express21, 9839–9850 (2013)..

Zhang, Y. et al. jGCaMP8 fast genetically encoded calcium indicators. (2020). athttps://doi.org/10.25378/janelia.13148243https://doi.org/10.25378/janelia.13148243.

Zeng, S. Q. et al. Simultaneous compensation for spatial and temporal dispersion of acousto-optical deflectors for two-dimensional scanning with a single prism.Opt. Lett.31, 1091–1093 (2006)..

Zeng, S. Q. et al. Analysis of the dispersion compensation of acousto-optic deflectors used for multiphoton imaging.J. Biomed. Opt.12, 024015 (2007)..

Chen, T. W. et al. Ultrasensitive fluorescent proteins for imaging neuronal activity.Nature499, 295–300 (2013)..

Sadagopan, S.&Wang, X. Q. Level invariant representation of sounds by populations of neurons in primary auditory cortex.J. Neurosci.28, 3415–3426 (2008)..

Rothschild, G., Nelken, I.&Mizrahi, A. Functional organization and population dynamics in the mouse primary auditory cortex.Nat. Neurosci.13, 353–360 (2010)..

Wong, A. B.&Borst, J. G. G. Tonotopic and non-auditory organization of the mouse dorsal inferior colliculus revealed by two-photon imaging.eLife8, e49091 (2019)..

Dupriez, P. Ultrafast fiber lasers for multiphoton microscopy.PhotonicsViews16, 70–73 (2019)..

Hellerer, T. et al. 920 nm fiber laser delivering 100 fs pulses for nonlinear microscopy. Proceedings of SPIE 10082, Multiphoton Microscopy in the Biomedical Sciences XIX. San Francisco, California, United States: SPIE, 2019, 108820U,https://doi.org/10.1117/12.2507753https://doi.org/10.1117/12.2507753.

Taberner, A. M.&Liberman, M. C. Response properties of single auditory nerve fibers in the mouse.J. Neurophysiol.93, 557–569 (2005)..

Otsu, Y. et al. Optical monitoring of neuronal activity at high frame rate with a digital random-access multiphoton (RAMP) microscope.J. Neurosci. Methods173, 259–270 (2008)..

Gao, L. X.&Wang, X. Q. Intracellular neuronal recording in Awake nonhuman primates.Nat. Protoc.15, 3615–3631 (2020)..

Dittgen, T. et al. Lentivirus-based genetic manipulations of cortical neurons and their optical and electrophysiological monitoring in vivo.Proc. Natl Acad. Sci. USA101, 18206–18211 (2004)..

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