Two-photon laser scanning microscopy with electrowetting-based prism scanning

Supekar, Omkar D.; Ozbay, Baris N.; Nystrom, Philip D.; Futia, Gregory L.; Restrepo, Diego; Gibson, Emily A.; Gopinath, Juliet T.; Bright, Victor M.

doi:10.1364/boe.8.005412

Cited by 39 publications

(22 citation statements)

References 32 publications

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“…And finally, we applied this method to several relevant in-vivo dynamically varying samples such as cells, muscle mitochondrial activity, intravital imaging of blood flow in the skull, and neural firing in the barrel cortex. Liquid lens technology has recently attracted attention for applications such as headmountable brain neural activity measurement [39], or endoscopic imaging by providing both lateral [40] and axial [41,42] scanning. Due to their high potential for low cost and stability, many different methods of producing liquid lenses have been proposed.…”

Section: Discussionmentioning

confidence: 99%

Five-dimensional two-photon volumetric microscopy of in-vivo dynamic activities using liquid lens remote focusing

Tehrani

Latchoumane

Southern

et al. 2019

Biomed. Opt. Express

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Multi-photon scanning microscopy provides a robust tool for optical sectioning, which can be used to capture fast biological events such as blood flow, mitochondrial activity, and neuronal action potentials. For many studies, it is important to visualize several different focal planes at a rate akin to the biological event frequency. Typically, a microscope is equipped with mechanical elements to move either the sample or the objective lens to capture volumetric information, but these strategies are limited due to their slow speeds or inertial artifacts. To overcome this problem, remote focusing methods have been developed to shift the focal plane axially without physical movement of the sample or the microscope. Among these methods is liquid lens technology, which adjusts the focus of the lens by changing the wettability of the liquid and hence its curvature. Liquid lenses are inexpensive active optical elements that have the potential for fast multi-photon volumetric imaging, hence a promising and accessible approach for the study of biological systems with complex dynamics. Although remote focusing using liquid lens technology can be used for volumetric point scanning multi-photon microscopy, optical aberrations and the effects of high energy laser pulses have been concerns in its implementation. In this paper, we characterize a liquid lens and validate its use in relevant biological applications. We measured optical aberrations that are caused by the liquid lens, and calculated its response time, defocus hysteresis, and thermal response to a pulsed laser. We applied this method of remote focusing for imaging and measurement of multiple in-vivo specimens, including mesenchymal stem cell dynamics, mouse tibialis anterior muscle mitochondrial electrical potential fluctuations, and mouse brain neural activity. Our system produces 5 dimensional (x,y,z,λ,t) data sets at the speed of 4.2 volumes per second over volumes as large as 160 x 160 x 35 µm 3 .

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Section: Discussionmentioning

confidence: 99%

Five-dimensional two-photon volumetric microscopy of in-vivo dynamic activities using liquid lens remote focusing

Tehrani

Latchoumane

Southern

et al. 2019

Biomed. Opt. Express

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“…The geometric configuration used in our simulation is based on our actual EWOD lenses and prisms [25,27,36,37]. The devices are constructed in cylindrical glass tubes with ITO sidewall electrodes patterned using a 3D printing-assisted shadow masking technique.…”

Section: Geometrical Configuration and Numerical Methodsmentioning

confidence: 99%

Numerical analysis of wavefront aberration correction using multielectrode electrowetting-based devices

et al. 2017

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We present numerical simulations of multielectrode electrowetting devices used in a novel optical design to correct wavefront aberration. Our optical system consists of two multielectrode devices, preceded by a single fixed lens. The multielectrode elements function as adaptive optical devices that can be used to correct aberrations inherent in many imaging setups, biological samples, and the atmosphere. We are able to accurately simulate the liquid-liquid interface shape using computational fluid dynamics. Ray tracing analysis of these surfaces shows clear evidence of aberration correction. To demonstrate the strength of our design, we studied three different input aberrations mixtures that include astigmatism, coma, trefoil, and additional higher order aberration terms, with amplitudes as large as one wave at 633 nm.

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“…The first adaptive prisms reported were based on electrowetting on dielectric. [24][25][26] However, the scanning angle range of electrowetting-based prisms is too narrow to achieve a large field of view. Recently, we introduced a piezo-actuated adaptive prism with a large tuning range 27,28 as a suitable alternative to galvanometer mirror that enables an in-line transmissive configuration with large scanning angle range.…”

Section: Introductionmentioning

confidence: 99%

3D-scanning microscopy with adaptive lenses and prisms for zebrafish studies

Wang

Lemke

Wapler

et al. 2021

J. Optical Microsystems

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Point-scanning-based microscopy systems require combination of axial and lateral scanning to obtain three-dimensional (3D) data. Axial scanning was commonly achieved by mechanical displacement of the objective or the sample. To improve, various adaptive lens-based solutions have been reported to circumvent the need for mechanically moving parts. The lateral scanning is predominantly implemented using galvanometric mirrors. Although the performance of such devices is flawless, they require bulky, folded beam-paths that make their incorporation in compact hand-held devices challenging. Recently, we introduced an adaptive prism as a transmissive device that enables lateral scanning. We demonstrate the first all-adaptive 3D scanning in laser scanning microscopes employing a compact in-line transmission geometry without mechanically moving parts and beam folding, combining an adaptive lens and a novel adaptive prism. Characterization of the all-adaptive microscope performance shows a lateral tuning range of approximately X ¼ Y ¼ 130 μm and an axial tuning range of about Z ¼ 500 μm. We successfully demonstrate 3D raster scanning of the fluorescence of a thyroid of a zebrafish embryo.

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Two-photon laser scanning microscopy with electrowetting-based prism scanning

Cited by 39 publications

References 32 publications

Five-dimensional two-photon volumetric microscopy of in-vivo dynamic activities using liquid lens remote focusing

Five-dimensional two-photon volumetric microscopy of in-vivo dynamic activities using liquid lens remote focusing

Numerical analysis of wavefront aberration correction using multielectrode electrowetting-based devices

3D-scanning microscopy with adaptive lenses and prisms for zebrafish studies

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