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PreprintCC BYbioRxiv

Measuring neuronal input-output operations in vivo with random access projection microscopy

  • M. E. Xie
  • M. Seyedolmohadesin
  • M. He
  • L. Li
  • Y. A. Hao
  • D. Flickinger
  • R. Gibiser
  • J. King
  • K. Driessen
  • J. Park
  • R. C. Peene
  • H. Loeffler
14 more authors
  • K. Nguyen
  • M. Tom
  • A. Williford
  • J. Swapp
  • G. Tononi
  • C. Cirelli
  • M. Z. Lin
  • B. E. Kimmel
  • J. Glaser
  • M. D. Golub
  • A. S. Charles
  • K. Svoboda
  • G. Jaindl
  • K. Podgorski1
  1. 1Allen Institute, Neural Dynamics, Seattle, WA
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Abstract

Neurons transform thousands of synaptic inputs into spiking output, and understanding this transformation requires measuring many submicron synapses at millisecond timescales in vivo. Here we present SLAP2, a two-photon random access projection microscope with no per-target access time cost, yielding over 5-fold faster imaging than existing methods, and performant real-time hardware motion correction. Flexible scanning modes enable measurement of hundreds of synapses at ~200 Hz, >800 synapses volumetrically at 50-100 Hz, and compartment-resolved dendritic voltage at 2-10 kHz in behaving mice. Simultaneous dendritic glutamate and somatic calcium imaging in visual cortex pyramidal neurons revealed nonlinear input-output amplification of stimulus onset and preferred stimulus orientation responses. Amplified features are associated with spatially organized inputs and stronger trial-to-trial coupling between synaptic activity and somatic output. These measurements establish SLAP2 as a powerful tool for recording intricate patterns of subcellular fluorescence dynamics in behaving animals.

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Cite

BibTeXRIS
BibTeX
@misc{xie2026measuring,
  title = {{Measuring neuronal input-output operations in vivo with random access projection microscopy}},
  author = {M. E. Xie and M. Seyedolmohadesin and M. He and L. Li and Y. A. Hao and D. Flickinger and R. Gibiser and J. King and K. Driessen and J. Park and R. C. Peene and H. Loeffler and K. Nguyen and M. Tom and A. Williford and J. Swapp and G. Tononi and C. Cirelli and M. Z. Lin and B. E. Kimmel and J. Glaser and M. D. Golub and A. S. Charles and K. Svoboda and G. Jaindl and K. Podgorski},
  howpublished = {bioRxiv},
  year = {2026},
  month = nov,
  doi = {10.64898/2026.11.05.764682},
  url = {https://www.biorxiv.org/content/10.64898/2026.11.05.764682v1},
  note = {CC BY},
}
RIS
TY  - UNPB
TI  - Measuring neuronal input-output operations in vivo with random access projection microscopy
AU  - M. E. Xie
AU  - M. Seyedolmohadesin
AU  - M. He
AU  - L. Li
AU  - Y. A. Hao
AU  - D. Flickinger
AU  - R. Gibiser
AU  - J. King
AU  - K. Driessen
AU  - J. Park
AU  - R. C. Peene
AU  - H. Loeffler
AU  - K. Nguyen
AU  - M. Tom
AU  - A. Williford
AU  - J. Swapp
AU  - G. Tononi
AU  - C. Cirelli
AU  - M. Z. Lin
AU  - B. E. Kimmel
AU  - J. Glaser
AU  - M. D. Golub
AU  - A. S. Charles
AU  - K. Svoboda
AU  - G. Jaindl
AU  - K. Podgorski
PY  - 2026
DA  - 2026/11/05
PB  - bioRxiv
DO  - 10.64898/2026.11.05.764682
UR  - https://www.biorxiv.org/content/10.64898/2026.11.05.764682v1
AB  - Neurons transform thousands of synaptic inputs into spiking output, and understanding this transformation requires measuring many submicron synapses at millisecond timescales in vivo. Here we present SLAP2, a two-photon random access projection microscope with no per-target access time cost, yielding over 5-fold faster imaging than existing methods, and performant real-time hardware motion correction. Flexible scanning modes enable measurement of hundreds of synapses at ~200 Hz, >800 synapses volumetrically at 50-100 Hz, and compartment-resolved dendritic voltage at 2-10 kHz in behaving mice. Simultaneous dendritic glutamate and somatic calcium imaging in visual cortex pyramidal neurons revealed nonlinear input-output amplification of stimulus onset and preferred stimulus orientation responses. Amplified features are associated with spatially organized inputs and stronger trial-to-trial coupling between synaptic activity and somatic output. These measurements establish SLAP2 as a powerful tool for recording intricate patterns of subcellular fluorescence dynamics in behaving animals.
ER  -