Rapid adaptive optical recovery of optimal resolution over large volumes

Wang, Kai; Milkie, Daniel E.; Saxena, Ankur; Engerer, Peter; Misgeld, Thomas; Bronner‐Fraser, Marianne; Mumm, Jeff S.; Betzig, Eric

doi:10.1038/nmeth.2925

Cited by 252 publications

(208 citation statements)

References 33 publications

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“…To overcome this problem, adaptive optics may be one of the potential solutions. [64][65][66] In addition, the design of°uorescent probes with higher quantum efciency, less photobleaching, and smaller sizes will notably improve the resolution. We expect that further development of super-resolution microscopy will yield e±cient and critical insights into biological science.…”

Section: Discussionmentioning

confidence: 99%

Super-resolution microscopy and its applications in neuroscience

Wang

Zhu

et al. 2017

J. Innov. Opt. Health Sci.

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Optical microscopy promises researchers to see most tiny substances directly. However, the resolution of conventional microscopy is restricted by the di®raction limit. This makes it a challenge to observe subcellular processes happened in nanoscale. The development of superresolution microscopy provides a solution to this challenge. Here, we brie°y review several commonly used super-resolution techniques, explicating their basic principles and applications in biological science, especially in neuroscience. In addition, characteristics and limitations of each technique are compared to provide a guidance for biologists to choose the most suitable tool.

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Section: Discussionmentioning

confidence: 99%

Super-resolution microscopy and its applications in neuroscience

Wang

Zhu

et al. 2017

J. Innov. Opt. Health Sci.

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“…Specifically, the SWIR beam, which can be focused deep inside the tissue, can be used as a reference point ("guiding star") allowing us to adjust the phase of the NIR beam to achieve the maximum spatial overlap of their focal volumes. An NIR beam has been recently introduced to correct distortions of the focal spot in the conventional (degenerate) 2-photon microscopy [48]. The same correction procedure will also compensate for chromatic aberrations of the objective allowing implementation of standard objectives used in degenerate 2-photon microscopy for nondegenerate excitation.…”

Section: Discussionmentioning

confidence: 99%

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy

Yang¹,

Abashin²,

Saisan³

et al. 2016

Opt. Express

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Non-degenerate 2-photon excitation (ND-2PE) of a fluorophore with two laser beams of different photon energies offers an independent degree of freedom in tuning of the photon flux for each beam. This feature takes advantage of the infrared wavelengths used in degenerate 3-photon excitation (D-3PE) microscopy to achieve increased penetration depths, while preserving a relatively high 2-photon excitation cross section in comparison to that of D-3PE. Here, using spatially and temporally aligned Ti:Sapphire laser and optical parametric oscillator beams operating at near infrared (NIR) and short-wavelength infrared (SWIR) optical frequencies, we employ ND-2PE and provide a practical demonstration that a constant fluorophore emission intensity is achievable deeper into a scattering medium using ND-2PE as compared to the commonly used degenerate 2-photon excitation (D-2PE). References and links1. K. Svoboda and R. Yasuda, "Principles of two-photon excitation microscopy and its applications to neuroscience," Neuron 50(6), 823-839 (2006). 2. J. N. Kerr and W. Denk, "Imaging in vivo: watching the brain in action," Nat. Rev. Neurosci. 9(3), 195-205 (2008). 3. W. Denk and K. Svoboda, "Photon upmanship: why multiphoton imaging is more than a gimmick," Neuron 18(3), 351-357 (1997). 4. F. Helmchen and W. Denk, "Deep tissue two-photon microscopy," Nat. Methods 2(12), 932-940 (2005 11. D. Kobat, N. G. Horton, and C. Xu, "In vivo two-photon microscopy to 1.6-mm depth in mouse cortex," J. Biomed. Opt. 16(10), 106014 (2011). 12. R. Kawakami, K. Sawada, A. Sato, T. Hibi, Y. Kozawa, S. Sato, H. Yokoyama, and T. Nemoto, "Visualizing hippocampal neurons with in vivo two-photon microscopy using a 1030 nm picosecond pulse laser," Sci. Rep. 3, 1014 (2013). 13. P. Theer and W. Denk, "On the fundamental imaging-depth limit in two-photon microscopy," J. Opt. Soc. Am.A adaptive optical recovery of optimal resolution over large volumes," Nat. Methods 11(6), 625-628 (2014).

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“…Of course, aberrations introduced by the sample itself also degrade image quality and it is those aberrations that are most difficult to correct at runtime due to their modal complexity, large amplitude, and high spatial variability. 26 Additionally, another problem inherent to light-sheet microscopy degrades image quality due to the uncoupled nature of excitation and detection in LSFM, the aberrations encountered in each arm are independent too and this can lead to a walkoff between the light sheet and the detection FOV when large samples are imaged at depths. Adaptive optics 27 provides a means to compensate for these aberrations and has recently been applied to improve LSFM.…”

Section: Discussionmentioning

confidence: 99%

Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains

et al. 2015

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Abstract. Comprehensive mapping and quantification of neuronal projections in the central nervous system requires high-throughput imaging of large volumes with microscopic resolution. To this end, we have developed a confocal light-sheet microscope that has been optimized for three-dimensional (3-D) imaging of structurally intact clarified whole-mount mouse brains. We describe the optical and electromechanical arrangement of the microscope and give details on the organization of the microscope management software. The software orchestrates all components of the microscope, coordinates critical timing and synchronization, and has been written in a versatile and modular structure using the LabVIEW language. It can easily be adapted and integrated to other microscope systems and has been made freely available to the light-sheet community. The tremendous amount of data routinely generated by light-sheet microscopy further requires novel strategies for data handling and storage. To complete the full imaging pipeline of our high-throughput microscope, we further elaborate on big data management from streaming of raw images up to stitching of 3-D datasets. The mesoscale neuroanatomy imaged at micron-scale resolution in those datasets allows characterization and quantification of neuronal projections in unsectioned mouse brains.

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Rapid adaptive optical recovery of optimal resolution over large volumes

Cited by 252 publications

References 33 publications

Super-resolution microscopy and its applications in neuroscience

Super-resolution microscopy and its applications in neuroscience

Non-degenerate 2-photon excitation in scattering medium for fluorescence microscopy

Comprehensive optical and data management infrastructure for high-throughput light-sheet microscopy of whole mouse brains

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