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A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity
We constructed a simple and compact imaging system designed specifically for the recording of fast neuronal activity in a 3D volume. The system uses an Yb:KYW femtosecond laser we designed for use with acousto-optic deflection. An integrated two-axis acousto-optic deflector, driven by digitally synt...
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2007
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1933593/ https://www.ncbi.nlm.nih.gov/pubmed/17684546 http://dx.doi.org/10.1371/journal.pone.0000699 |
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author | Vučinić, Dejan Sejnowski, Terrence J. |
author_facet | Vučinić, Dejan Sejnowski, Terrence J. |
author_sort | Vučinić, Dejan |
collection | PubMed |
description | We constructed a simple and compact imaging system designed specifically for the recording of fast neuronal activity in a 3D volume. The system uses an Yb:KYW femtosecond laser we designed for use with acousto-optic deflection. An integrated two-axis acousto-optic deflector, driven by digitally synthesized signals, can target locations in three dimensions. Data acquisition and the control of scanning are performed by a LeCroy digital oscilloscope. The total cost of construction was one order of magnitude lower than that of a typical Ti:sapphire system. The entire imaging apparatus, including the laser, fits comfortably onto a small rig for electrophysiology. Despite the low cost and simplicity, the convergence of several new technologies allowed us to achieve the following capabilities: i) full-frame acquisition at video rates suitable for patch clamping; ii) random access in under ten microseconds with dwelling ability in the nominal focal plane; iii) three-dimensional random access with the ability to perform fast volume sweeps at kilohertz rates; and iv) fluorescence lifetime imaging. We demonstrate the ability to record action potentials with high temporal resolution using intracellularly loaded potentiometric dye di-2-ANEPEQ. Our design proffers easy integration with electrophysiology and promises a more widespread adoption of functional two-photon imaging as a tool for the study of neuronal activity. The software and firmware we developed is available for download at http://neurospy.org/ under an open source license. |
format | Text |
id | pubmed-1933593 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-19335932007-08-08 A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity Vučinić, Dejan Sejnowski, Terrence J. PLoS One Research Article We constructed a simple and compact imaging system designed specifically for the recording of fast neuronal activity in a 3D volume. The system uses an Yb:KYW femtosecond laser we designed for use with acousto-optic deflection. An integrated two-axis acousto-optic deflector, driven by digitally synthesized signals, can target locations in three dimensions. Data acquisition and the control of scanning are performed by a LeCroy digital oscilloscope. The total cost of construction was one order of magnitude lower than that of a typical Ti:sapphire system. The entire imaging apparatus, including the laser, fits comfortably onto a small rig for electrophysiology. Despite the low cost and simplicity, the convergence of several new technologies allowed us to achieve the following capabilities: i) full-frame acquisition at video rates suitable for patch clamping; ii) random access in under ten microseconds with dwelling ability in the nominal focal plane; iii) three-dimensional random access with the ability to perform fast volume sweeps at kilohertz rates; and iv) fluorescence lifetime imaging. We demonstrate the ability to record action potentials with high temporal resolution using intracellularly loaded potentiometric dye di-2-ANEPEQ. Our design proffers easy integration with electrophysiology and promises a more widespread adoption of functional two-photon imaging as a tool for the study of neuronal activity. The software and firmware we developed is available for download at http://neurospy.org/ under an open source license. Public Library of Science 2007-08-08 /pmc/articles/PMC1933593/ /pubmed/17684546 http://dx.doi.org/10.1371/journal.pone.0000699 Text en Vucinic, Sejnowski. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Vučinić, Dejan Sejnowski, Terrence J. A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity |
title | A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity |
title_full | A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity |
title_fullStr | A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity |
title_full_unstemmed | A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity |
title_short | A Compact Multiphoton 3D Imaging System for Recording Fast Neuronal Activity |
title_sort | compact multiphoton 3d imaging system for recording fast neuronal activity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1933593/ https://www.ncbi.nlm.nih.gov/pubmed/17684546 http://dx.doi.org/10.1371/journal.pone.0000699 |
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