Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain
1300 nm three-photon calcium imaging has emerged as a useful technique to allow calcium imaging in deep brain regions. Application to large-scale neural activity imaging entails a careful balance between recording fidelity and perturbation to the sample. We calculated and experimentally verified the...
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eLife Sciences Publications Ltd
2020-01-01
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Online Access: | https://elifesciences.org/articles/53205 |
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author | Tianyu Wang Chunyan Wu Dimitre G Ouzounov Wenchao Gu Fei Xia Minsu Kim Xusan Yang Melissa R Warden Chris Xu |
author_facet | Tianyu Wang Chunyan Wu Dimitre G Ouzounov Wenchao Gu Fei Xia Minsu Kim Xusan Yang Melissa R Warden Chris Xu |
author_sort | Tianyu Wang |
collection | DOAJ |
description | 1300 nm three-photon calcium imaging has emerged as a useful technique to allow calcium imaging in deep brain regions. Application to large-scale neural activity imaging entails a careful balance between recording fidelity and perturbation to the sample. We calculated and experimentally verified the excitation pulse energy to achieve the minimum photon count required for the detection of calcium transients in GCaMP6s-expressing neurons for 920 nm two-photon and 1320 nm three-photon excitation. By considering the combined effects of in-focus signal attenuation and out-of-focus background generation, we quantified the cross-over depth beyond which three-photon microscopy outpeforms two-photon microscopy in recording fidelity. Brain tissue heating by continuous three-photon imaging was simulated with Monte Carlo method and experimentally validated with immunohistochemistry. Increased immunoreactivity was observed with 150 mW excitation power at 1 and 1.2 mm imaging depths. Our analysis presents a translatable model for the optimization of three-photon calcium imaging based on experimentally tractable parameters. |
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issn | 2050-084X |
language | English |
last_indexed | 2024-04-12T16:43:36Z |
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spelling | doaj.art-bb1f5a298fca4712ae992c821a4f0f792022-12-22T03:24:41ZengeLife Sciences Publications LtdeLife2050-084X2020-01-01910.7554/eLife.53205Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brainTianyu Wang0https://orcid.org/0000-0002-6087-6376Chunyan Wu1https://orcid.org/0000-0002-6294-4512Dimitre G Ouzounov2Wenchao Gu3Fei Xia4https://orcid.org/0000-0001-6591-8769Minsu Kim5Xusan Yang6Melissa R Warden7https://orcid.org/0000-0003-2240-3997Chris Xu8https://orcid.org/0000-0002-3493-6427School of Applied and Engineering Physics, Cornell University, Ithaca, United StatesSchool of Applied and Engineering Physics, Cornell University, Ithaca, United States; College of Veterinary Medicine, Cornell University, Ithaca, United StatesSchool of Applied and Engineering Physics, Cornell University, Ithaca, United StatesDepartment of Neurobiology and Behavior, Cornell University, Ithaca, United StatesMeining School of Biomedical Engineering, Cornell University, Ithaca, United StatesCollege of Human Ecology, Cornell University, Ithaca, United StatesSchool of Applied and Engineering Physics, Cornell University, Ithaca, United StatesDepartment of Neurobiology and Behavior, Cornell University, Ithaca, United StatesSchool of Applied and Engineering Physics, Cornell University, Ithaca, United States1300 nm three-photon calcium imaging has emerged as a useful technique to allow calcium imaging in deep brain regions. Application to large-scale neural activity imaging entails a careful balance between recording fidelity and perturbation to the sample. We calculated and experimentally verified the excitation pulse energy to achieve the minimum photon count required for the detection of calcium transients in GCaMP6s-expressing neurons for 920 nm two-photon and 1320 nm three-photon excitation. By considering the combined effects of in-focus signal attenuation and out-of-focus background generation, we quantified the cross-over depth beyond which three-photon microscopy outpeforms two-photon microscopy in recording fidelity. Brain tissue heating by continuous three-photon imaging was simulated with Monte Carlo method and experimentally validated with immunohistochemistry. Increased immunoreactivity was observed with 150 mW excitation power at 1 and 1.2 mm imaging depths. Our analysis presents a translatable model for the optimization of three-photon calcium imaging based on experimentally tractable parameters.https://elifesciences.org/articles/53205calciumthree-photon3-photontwo-photon2-photonneural imaging |
spellingShingle | Tianyu Wang Chunyan Wu Dimitre G Ouzounov Wenchao Gu Fei Xia Minsu Kim Xusan Yang Melissa R Warden Chris Xu Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain eLife calcium three-photon 3-photon two-photon 2-photon neural imaging |
title | Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain |
title_full | Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain |
title_fullStr | Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain |
title_full_unstemmed | Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain |
title_short | Quantitative analysis of 1300-nm three-photon calcium imaging in the mouse brain |
title_sort | quantitative analysis of 1300 nm three photon calcium imaging in the mouse brain |
topic | calcium three-photon 3-photon two-photon 2-photon neural imaging |
url | https://elifesciences.org/articles/53205 |
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