High-speed microscopy with an electrically tunable lens to image the dynamics of <i>in vivo</i> molecular complexes

Nakai, Yuichiro; Ozeki, Mitsunori; Hiraiwa, Takumi; Tanimoto, Ryuichi; Funahashi, Akira; Hiroi, Noriko; Taniguchi, Atsushi; Nonaka, Shigenori; Boilot, Viviane; Shrestha, Roshan L.; Clark, Joanna; Tamura, Naoka; Draviam, Viji M.; Oku, Hiromasa

doi:10.1063/1.4905330

Cited by 45 publications

(32 citation statements)

References 27 publications

(28 reference statements)

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“…This results in the aforementioned artefact in the first few lines of each frame and is caused by fast oscillations in the ETL’s focal power after a rapid change in driving current (see link to specifications, Table 1). The stabilization time of ∼10 ms (for travel distances of ∼50 µm) we observed is consistent with earlier reports 25,61 . For simplicity, we here used scans with more lines, such that the artefact was outside the IPL.…”

Section: Discussionsupporting

confidence: 93%

The temporal structure of the inner retina at a single glance

Zhao

Klindt

Chagas

et al. 2019

Preprint

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21The retina decomposes visual stimuli into parallel channels that encode different features of the visual 22 environment. Central to this computation is the synaptic processing in a dense and thick layer of 23 neuropil, the so-called inner plexiform layer (IPL). Here, different types of bipolar cells stratifying at 24 distinct depths relay the excitatory feedforward drive from photoreceptors to amacrine and ganglion 25 cells. Current experimental techniques for studying processing in the IPL do not allow imaging the entire 26 IPL simultaneously in the intact tissue. Here, we extend a two-photon microscope with an electrically 27 tunable lens allowing us to obtain optical vertical slices of the IPL, which provide a complete picture of 28 the response diversity of bipolar cells at a "single glance". The nature of these axial recordings 29additionally allowed us to isolate and investigate batch effects, i.e. inter-experimental variations 30 resulting in systematic differences in response speed. As a proof of principle, we developed a simple 31 model that disentangles biological from experimental causes of variability, and allowed us to recover 32 the characteristic gradient of response speeds across the IPL with higher precision than before. Our 33 new framework will make it possible to study the computations performed in the central synaptic layer 34 of the retina more efficiently. 353

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Section: Discussionsupporting

confidence: 93%

The temporal structure of the inner retina at a single glance

Zhao

Klindt

Chagas

et al. 2019

Preprint

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“…Partly in response to this specific demand, in recent years several technologies have been developed to tune the optical power of lenses (Berge, 2005;Ren et al, 2006;Bernet et al, 2013) or Micro-Electro-Mechanical System mirrors (Qi et al, 2004;Shain et al, 2017). Some of these systems, like the tunable polymer-filled lenses (Ren et al, 2006;Jabbour et al, 2014;Nakai et al, 2015) or lenses based on electrowetting (Berge, 2005) are now available. However, these systems possess a minimal response time of more than several milliseconds, which is inadequate for high-speed recording.…”

Section: Introductionmentioning

confidence: 99%

Remote focusing in confocal microscopy by means of a modified Alvarez lens

Bawart

Jesacher

Bernet

et al. 2018

Journal of Microscopy

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Alvarez lenses are actuated lens-pairs which allow one to tune the optical power by mechanical displacement of subelements. Here, we show that a recently realized modified Alvarez lens design which does not require mechanical actuation can be integrated into a confocal microscope. Instead of mechanically moving them, the sublenses are imaged onto each other in a 4f-configuration, where the lateral image shift leading to a change in optical power is created by a galvo-mirror. The avoidance of mechanical lens shifts leads to a large speed gain for axial (and hence also 3D) image scans compared to classical Alvarez lenses. We demonstrate that the suggested operation principle is compatible with confocal microscopy. In order to optimize the system, we have drawn advantage of the flexibility a liquid-crystal spatial light modulator offers for the implementation. For given specifications, dedicated diffractive optical elements or freeform elements can be used in combination with resonant galvo-scanners or acousto-optic beam deflectors, to achieve even faster z-scans than reported here, reaching video rate.

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“…17 Tunable lenses offer the advantage of a large focal range without any moving parts within a few milliseconds only, thus allowing a compact setup and fast switching among different foci. Hence, they have been used in many optical systems, especially in microscopes, to make sure that the moving object stays in focus 18,19 or to focus on different optical plans in z-directions. 20,21 On the other hand, mimicking of the eye's pupil shift (e.g., due to head motion) can be realized by a laterally moving aperture on an x − y stage or by a spatial light modulator.…”

Section: Introductionmentioning

confidence: 99%

Focus and perspective adaptive digital surgical microscope: optomechanical design and experimental implementation

2017

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This paper relates to the improvement of conventional surgical stereo microscopy via the application of digital recording devices and adaptive optics. The research is aimed at improving the working conditions of the surgeon during the operation, such that free head movement is possible. The depth clues known from conventional stereo microscopy in interaction with the human eye’s functionality, such as convergence, disparity, angular elevation, parallax, and accommodation, are implemented in a digital recording system via adaptive optomechanical components. Two laterally moving pupil apertures have been used mimicking the digital implementation of the eye’s vergence and head motion. The natural eye’s accommodation is mimicked via the application of a tunable lens. Additionally, another system has been built, which enables tracking the surgeon’s eye pupil through a digital displaying stereoscopic microscope to supply the necessary information for steering the recording system. The optomechanical design and experimental results for both systems, digital recording stereoscopic microscope and pupil tracking system, are shown.

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High-speed microscopy with an electrically tunable lens to image the dynamics of in vivo molecular complexes

Cited by 45 publications

References 27 publications

The temporal structure of the inner retina at a single glance

The temporal structure of the inner retina at a single glance

Remote focusing in confocal microscopy by means of a modified Alvarez lens

Focus and perspective adaptive digital surgical microscope: optomechanical design and experimental implementation

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