Two-dimensional electro-optical multiphoton microscopy
The development of genetically encoded fluorescent indicators of neural activity with millisecond dynamics has generated demand for ever faster two-photon (2P) imaging systems, but acoustic and mechanical beam scanning technologies are approaching fundamental limits. We demonstrate that potassium ta...
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| Published in | Neurophotonics (Print) Vol. 11; no. 2; p. 025005 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
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Society of Photo-Optical Instrumentation Engineers
01.04.2024
SPIE |
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| Abstract | The development of genetically encoded fluorescent indicators of neural activity with millisecond dynamics has generated demand for ever faster two-photon (2P) imaging systems, but acoustic and mechanical beam scanning technologies are approaching fundamental limits. We demonstrate that potassium tantalate niobate (KTN) electro-optical deflectors (EODs), which are not subject to the same fundamental limits, are capable of ultrafast two-dimensional (2D) 2P imaging
.
To determine if KTN-EODs are suitable for 2P imaging, compatible with 2D scanning, and capable of ultrafast
imaging of genetically encoded indicators with millisecond dynamics.
The performance of a commercially available KTN-EOD was characterized across a range of drive frequencies and laser parameters relevant to
2P microscopy. A second KTN-EOD was incorporated into a dual-axis scan module, and the system was validated by imaging signals
from ASAP3, a genetically encoded voltage indicator.
Optimal KTN-EOD deflection of laser light with a central wavelength of 960 nm was obtained up to the highest average powers and pulse intensities tested (power: 350 mW; pulse duration: 118 fs). Up to 32 resolvable spots per line at a 560 kHz line scan rate could be obtained with single-axis deflection. The complete dual-axis EO 2P microscope was capable of imaging a
by
field-of-view at over 10 kHz frame rate with
lateral resolution. We demonstrate
imaging of neurons expressing ASAP3 with high temporal resolution.
We demonstrate the suitability of KTN-EODs for ultrafast 2P cellular imaging
, providing a foundation for future high-performance microscopes to incorporate emerging advances in KTN-based scanning technology. |
|---|---|
| AbstractList | The development of genetically encoded fluorescent indicators of neural activity with millisecond dynamics has generated demand for ever faster two-photon (2P) imaging systems, but acoustic and mechanical beam scanning technologies are approaching fundamental limits. We demonstrate that potassium tantalate niobate (KTN) electro-optical deflectors (EODs), which are not subject to the same fundamental limits, are capable of ultrafast two-dimensional (2D) 2P imaging
.
To determine if KTN-EODs are suitable for 2P imaging, compatible with 2D scanning, and capable of ultrafast
imaging of genetically encoded indicators with millisecond dynamics.
The performance of a commercially available KTN-EOD was characterized across a range of drive frequencies and laser parameters relevant to
2P microscopy. A second KTN-EOD was incorporated into a dual-axis scan module, and the system was validated by imaging signals
from ASAP3, a genetically encoded voltage indicator.
Optimal KTN-EOD deflection of laser light with a central wavelength of 960 nm was obtained up to the highest average powers and pulse intensities tested (power: 350 mW; pulse duration: 118 fs). Up to 32 resolvable spots per line at a 560 kHz line scan rate could be obtained with single-axis deflection. The complete dual-axis EO 2P microscope was capable of imaging a
by
field-of-view at over 10 kHz frame rate with
lateral resolution. We demonstrate
imaging of neurons expressing ASAP3 with high temporal resolution.
We demonstrate the suitability of KTN-EODs for ultrafast 2P cellular imaging
, providing a foundation for future high-performance microscopes to incorporate emerging advances in KTN-based scanning technology. The development of genetically encoded fluorescent indicators of neural activity with millisecond dynamics has generated demand for ever faster two-photon (2P) imaging systems, but acoustic and mechanical beam scanning technologies are approaching fundamental limits. We demonstrate that potassium tantalate niobate (KTN) electro-optical deflectors (EODs), which are not subject to the same fundamental limits, are capable of ultrafast two-dimensional (2D) 2P imaging in vivo.SignificanceThe development of genetically encoded fluorescent indicators of neural activity with millisecond dynamics has generated demand for ever faster two-photon (2P) imaging systems, but acoustic and mechanical beam scanning technologies are approaching fundamental limits. We demonstrate that potassium tantalate niobate (KTN) electro-optical deflectors (EODs), which are not subject to the same fundamental limits, are capable of ultrafast two-dimensional (2D) 2P imaging in vivo.To determine if KTN-EODs are suitable for 2P imaging, compatible with 2D scanning, and capable of ultrafast in vivo imaging of genetically encoded indicators with millisecond dynamics.AimTo determine if KTN-EODs are suitable for 2P imaging, compatible with 2D scanning, and capable of ultrafast in vivo imaging of genetically encoded indicators with millisecond dynamics.The performance of a commercially available KTN-EOD was characterized across a range of drive frequencies and laser parameters relevant to in vivo 2P microscopy. A second KTN-EOD was incorporated into a dual-axis scan module, and the system was validated by imaging signals in vivo from ASAP3, a genetically encoded voltage indicator.ApproachThe performance of a commercially available KTN-EOD was characterized across a range of drive frequencies and laser parameters relevant to in vivo 2P microscopy. A second KTN-EOD was incorporated into a dual-axis scan module, and the system was validated by imaging signals in vivo from ASAP3, a genetically encoded voltage indicator.Optimal KTN-EOD deflection of laser light with a central wavelength of 960 nm was obtained up to the highest average powers and pulse intensities tested (power: 350 mW; pulse duration: 118 fs). Up to 32 resolvable spots per line at a 560 kHz line scan rate could be obtained with single-axis deflection. The complete dual-axis EO 2P microscope was capable of imaging a 13 μm by 13 μm field-of-view at over 10 kHz frame rate with ∼0.5 μm lateral resolution. We demonstrate in vivo imaging of neurons expressing ASAP3 with high temporal resolution.ResultsOptimal KTN-EOD deflection of laser light with a central wavelength of 960 nm was obtained up to the highest average powers and pulse intensities tested (power: 350 mW; pulse duration: 118 fs). Up to 32 resolvable spots per line at a 560 kHz line scan rate could be obtained with single-axis deflection. The complete dual-axis EO 2P microscope was capable of imaging a 13 μm by 13 μm field-of-view at over 10 kHz frame rate with ∼0.5 μm lateral resolution. We demonstrate in vivo imaging of neurons expressing ASAP3 with high temporal resolution.We demonstrate the suitability of KTN-EODs for ultrafast 2P cellular imaging in vivo, providing a foundation for future high-performance microscopes to incorporate emerging advances in KTN-based scanning technology.ConclusionsWe demonstrate the suitability of KTN-EODs for ultrafast 2P cellular imaging in vivo, providing a foundation for future high-performance microscopes to incorporate emerging advances in KTN-based scanning technology. |
| Author | Stanek, Samuel Leger, James Newman, Zachary L. Farinella, Deano M. Gable, Jacob Kerlin, Aaron Jayakumar, Harishankar |
| Author_xml | – sequence: 1 givenname: Deano M. orcidid: 0000-0003-0312-0801 surname: Farinella fullname: Farinella, Deano M. email: farin012@umn.edu organization: University of Minnesota, Department of Neuroscience, Minneapolis, Minnesota, United States – sequence: 2 givenname: Samuel surname: Stanek fullname: Stanek, Samuel email: stane118@umn.edu organization: University of Minnesota, Department of Electrical and Computer Engineering, Minneapolis, Minnesota, United States – sequence: 3 givenname: Harishankar surname: Jayakumar fullname: Jayakumar, Harishankar email: harish@umn.edu organization: University of Minnesota, Department of Neuroscience, Minneapolis, Minnesota, United States – sequence: 4 givenname: Zachary L. orcidid: 0000-0002-4336-6585 surname: Newman fullname: Newman, Zachary L. email: newmanza@umn.edu organization: University of Minnesota, Department of Neuroscience, Minneapolis, Minnesota, United States – sequence: 5 givenname: Jacob surname: Gable fullname: Gable, Jacob email: gable098@umn.edu organization: University of Minnesota, Department of Neuroscience, Minneapolis, Minnesota, United States – sequence: 6 givenname: James orcidid: 0000-0001-9319-7196 surname: Leger fullname: Leger, James email: leger@umn.edu organization: University of Minnesota, Department of Electrical and Computer Engineering, Minneapolis, Minnesota, United States – sequence: 7 givenname: Aaron surname: Kerlin fullname: Kerlin, Aaron email: akerlin@umn.edu organization: University of Minnesota, Department of Neuroscience, Minneapolis, Minnesota, United States |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/38841422$$D View this record in MEDLINE/PubMed |
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