dandiarchive-dataset/s3://dandiarchive/dandisets/000454/0.230302.2331/dandiset.yaml

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  approach:
  - name: microscopy approach; cell population imaging
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  dataStandard:
  - identifier: RRID:SCR_015242
    name: Neurodata Without Borders (NWB)
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  measurementTechnique:
  - name: surgical technique
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  - name: two-photon microscopy technique
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  species:
  - identifier: http://purl.obolibrary.org/obo/NCBITaxon_10090
    name: Mus musculus - House mouse
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  variableMeasured:
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citation: Klienfeld, David; Yao, Pantong; Liu, Rui; Broginni, Thomas; Thunemann, Martin;
  University of California, San Diego; Boston University; University of California,
  San Diego (2023) Guide to the construction and use of an adaptive optics two-photon
  microscope with direct wavefront sensing (Version 0.230302.2331) [Data set]. DANDI
  archive. https://dandiarchive.org/dandiset/000454/0.230302.2331
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  email: dk@physics.ucsd.edu
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  name: Klienfeld, David
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  email: p1yao@health.ucsd.edu
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  name: Yao, Pantong
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  email: rul087@physics.ucsd.edu
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  name: Liu, Rui
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  name: Broginni, Thomas
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  name: Thunemann, Martin
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  url: http://www.bu.edu/
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  name: University of California, San Diego
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  name: Boston University
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  name: University of California, San Diego
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dateCreated: '2023-03-02T21:52:11.686223+00:00'
datePublished: '2023-03-02T23:31:53.735293+00:00'
description: Two-photon microscopy, combined with appropriate optical labeling, has
  enabled the study of structure and function throughout animals and their organ systems,
  especially nervous systems. This methodology enables, for example, the measurement
  and tracking of sub-micrometer structures within brain cells, the spatio-temporal
  mapping of spikes in individual neurons, and the spatio-temporal mapping of transmitter
  release in individual synapses. Yet the spatial resolution of two-photon microscopy
  rapidly degrades as imaging is attempted at depths more than a few scattering lengths
  into tissue, i.e., below the superficial layers that constitute the top 300 to 400 µm
  of neocortex. To obviate this limitation, we measure the wavefront of the guide
  star at the focus of the excitation beam and utilize adaptive optics that alters
  the incident wavefront to achieve an improved focal volume. We describe the construction,
  calibration, and operation of a two-photon microscope that incorporates adaptive
  optics to restore diffraction-limited resolution throughout the nearly 900 µm depth
  of mouse cortex. Our realization utilizes a guide star formed by excitation of red-shifted
  dye within the blood serum to directly measure the wavefront. We incorporate predominantly
  commercial optical, optomechanical, mechanical, and electronic components; computer
  aided design models of the exceptional custom components are supplied. The resultant
  adaptive-optics two-photon microscope is modular and allows for expanded imaging
  and optical excitation capabilities. We demonstrate our methodology in mouse neocortex
  by imaging the morphology of somatostatin-expressing neurons that lie 700 µm beneath
  the pia, calcium dynamics of layer 5b projection neurons, and thalamocortical glutamate
  transmission to L4 neurons.
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id: DANDI:000454/0.230302.2331
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name: Guide to the construction and use of an adaptive optics two-photon microscope
  with direct wavefront sensing
protocol: []
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