Enhanced volumetric imaging in 2‐photon microscopy via acoustic lens beam shaping

Piazza, Simonluca; Bianchini, Paolo; Sheppard, Colin J. R.; Diaspro, Alberto; Duocastella, Martí

doi:10.1002/jbio.201700050

Cited by 38 publications

(35 citation statements)

References 47 publications

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“…The new microscope opens the door to monitoring rapidly evolving processes or events that occur over temporally or spatially varying scales. We anticipate that coupling the technique with parallelized detection methods [9] or nonlinear excitation [15,29] will lead to improved signal-to-noise ratio, spatial resolution or penetration depth, helping to reconstruct complex 3D phenomena with maximal detail.…”

Section: Discussionmentioning

confidence: 99%

“…60 % for 40x objective and 45% for 20x objective lens. This problem could have been solved by placing the TAG lens before the scan head and expanding the beam diameter [15,29], but in our current commercial microscope this was not possible. The magnification factor caused by the difference of focal length between the relay lenses caused a smaller field of view, with the pixel size calibrated accordingly.…”

Section: Implementation Of the Volumetric Lissajous Confocal Microscopementioning

confidence: 99%

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Volumetric Lissajous Confocal Microscopy

Deguchi

Bianchini

Palazzolo

et al. 2019

Preprint

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Dynamic biological systems present challenges to existing three-dimensional (3D) optical microscopes because of their continuous temporal and spatial changes. Most techniques are based on rigid architectures, as in confocal microscopy, where a laser beam is sequentially scanned at a predefined spatial sampling rate and pixel dwell time. Here, we developed volumetric Lissajous confocal microscopy to achieve unsurpassed 3D scanning speed with a tunable sampling rate. The system combines an acoustic liquid lens for continuous axial focus translation with a resonant scanning mirror. Accordingly, the excitation beam follows a dynamic Lissajous trajectory enabling sub-millisecond acquisitions of image series containing 3D information at a sub-Nyquist sampling rate. By temporal accumulation and/or advanced interpolation algorithms, volumetric imaging rate is selectable using a post-processing step at the desired spatiotemporal resolution for events of interest. We demonstrate multicolor and calcium imaging over volumes of tens of cubic microns with acquisition speeds up to 5 kHz.

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Section: Discussionmentioning

confidence: 99%

Section: Implementation Of the Volumetric Lissajous Confocal Microscopementioning

confidence: 99%

Volumetric Lissajous Confocal Microscopy

Deguchi

Bianchini

Palazzolo

et al. 2019

Preprint

Self Cite

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“…3e). In contrast to PSF engineering techniques involving a non-diffracting illumination beam, e.g., using an axicon or tunable acoustic gradient-index lens 5,33 , this method is scalable in the DOF, allowing one to tailor different experimental specifications by tuning the refractive index without a dedicated optical system alignment.…”

Section: Clam Performancementioning

confidence: 99%

Parallelized volumetric fluorescence microscopy with a reconfigurable coded incoherent light-sheet array

Ren

Lai

et al. 2020

Light Sci Appl

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Parallelized fluorescence imaging has been a long-standing pursuit that can address the unmet need for a comprehensive three-dimensional (3D) visualization of dynamical biological processes with minimal photodamage. However, the available approaches are limited to incomplete parallelization in only two dimensions or sparse sampling in three dimensions. We hereby develop a novel fluorescence imaging approach, called coded light-sheet array microscopy (CLAM), which allows complete parallelized 3D imaging without mechanical scanning. Harnessing the concept of an "infinity mirror", CLAM generates a light-sheet array with controllable sheet density and degree of coherence. Thus, CLAM circumvents the common complications of multiple coherent light-sheet generation in terms of dedicated wavefront engineering and mechanical dithering/scanning. Moreover, the encoding of multiplexed optical sections in CLAM allows the synchronous capture of all sectioned images within the imaged volume. We demonstrate the utility of CLAM in different imaging scenarios, including a light-scattering medium, an optically cleared tissue, and microparticles in fluidic flow. CLAM can maximize the signal-to-noise ratio and the spatial duty cycle, and also provides a further reduction in photobleaching compared to the major scanning-based 3D imaging systems. The flexible implementation of CLAM regarding both hardware and software ensures compatibility with any light-sheet imaging modality and could thus be instrumental in a multitude of areas in biological research.

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“…[19,20] For a rectangular cavity, driving each piezoelectric pair with p different frequencies leads to a sinusoidally varying temporal and spatial refractive index that can be expressed as (see Figure S1, Supporting Information) [19,20] For a rectangular cavity, driving each piezoelectric pair with p different frequencies leads to a sinusoidally varying temporal and spatial refractive index that can be expressed as (see Figure S1, Supporting Information) …”

Section: Fast Acoustic Light Sculpting For On-demand Maskless Lithogrmentioning

confidence: 99%

Fast Acoustic Light Sculpting for On‐Demand Maskless Lithography

Surdo

2019

Advanced Science

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Light interference is the primary enabler of a number of optical maskless techniques for the large‐scale processing of materials at the nanoscale. However, methods controlling interference phenomena can be limited in speed, ease of implementation, or the selection of pattern designs. Here, an optofluidic system that employs acoustic standing waves in a liquid to produce complex interference patterns at sub‐microsecond temporal resolution, faster than the pulse‐to‐pulse period of many commercial laser systems, is presented. By controlling the frequency of the acoustic waves and the motion of a translation stage, additive and subtractive direct‐writing of tailored patterns over cm 2 areas with sub‐wavelength uniformity in periodicity and scalable spatial resolution, down to the nanometric range, are demonstrated. Such on‐the‐fly dynamic control of light enhances throughput and design flexibility of optical maskless lithography, helping to expand its application portfolio to areas as important as plasmonics, electronics, or metamaterials.

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Enhanced volumetric imaging in 2‐photon microscopy via acoustic lens beam shaping

Cited by 38 publications

References 47 publications

Volumetric Lissajous Confocal Microscopy

Volumetric Lissajous Confocal Microscopy

Parallelized volumetric fluorescence microscopy with a reconfigurable coded incoherent light-sheet array

Fast Acoustic Light Sculpting for On‐Demand Maskless Lithography

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