Field of view advantage of conjugate adaptive optics in microscopy applications

Mertz, Jérôme; Paudel, Hari P.; Bifano, Thomas G.

doi:10.1364/ao.54.003498

Cited by 94 publications

(98 citation statements)

References 28 publications

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“…More recently, this same principle of conjugate AO has made its way to the microscopy community. Reports in both simulation [6][7][8] and experiment [9][10][11][12][13] have demonstrated the FOV advantage of conjugate AO as well as its feasibility. For example, conjugate AO has been applied in scanning microscopy configurations using both one- 9 and twophoton 10,11 microscopy.…”

Section: Introductionmentioning

confidence: 92%

“…It has also been applied in widefield (i.e., nonscanning) microscopy configurations with transillumination geometries. 12,13 The last of these involved the use of an extended-source wavefront sensor where the transillumination itself served to determine the wavefront aberrations, obviating the need for a "guide star" within the sample. While in principle this same wavefront sensor could be applied in a fluorescence microscopy application, making use of the sample fluorescence itself as the illumination source to measure wavefront aberrations, we found this difficult to implement in practice.…”

Section: Introductionmentioning

confidence: 99%

“…3. Unlike pupil AO, which typically provides only a limited correction FOV in the presence of spatially varying aberrations, 12 conjugate AO is able to correct over almost the entire image FOV (here ∼400 μm × 400 μm), limited only by the size of the DM itself. Moreover, our OBM sensor is able to measure wavefronts at almost half video rate (requiring four frames, each at 40 Hz rate), enabling the conjugate AO to be performed in real time.…”

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confidence: 99%

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Widefield fluorescence microscopy with sensor-based conjugate adaptive optics using oblique back illumination

Bifano

Mertz

2016

J. Biomed. Opt.

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fluorescence microscopy with sensor-based conjugate adaptive optics using oblique back illumination," J. Abstract. We describe a wavefront sensor strategy for the implementation of adaptive optics (AO) in microscope applications involving thick, scattering media. The strategy is based on the exploitation of multiple scattering to provide oblique back illumination of the wavefront-sensor focal plane, enabling a simple and direct measurement of the flux-density tilt angles caused by aberrations at this plane. Advantages of the sensor are that it provides a large measurement field of view (FOV) while requiring no guide star, making it particularly adapted to a type of AO called conjugate AO, which provides a large correction FOV in cases when sample-induced aberrations arise from a single dominant plane (e.g., the sample surface). We apply conjugate AO here to widefield (i.e., nonscanning) fluorescence microscopy for the first time and demonstrate dynamic wavefront correction in a closed-loop implementation.

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Section: Introductionmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 99%

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confidence: 99%

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Widefield fluorescence microscopy with sensor-based conjugate adaptive optics using oblique back illumination

Bifano

Mertz

2016

J. Biomed. Opt.

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“…The volume wavefront aberrations were measured from different directions using multiple wavefront sensors. 121 However, although simulations have shown that microscopy will also benefit from multiconjugate adaptive optics, [122][123][124] directly applying this approach to microscopy has proven to be difficult, despite the improvements it would provide. This is primarily because it is not straightforward to separate aberrations from different layers in highly turbid media.…”

Section: Enlarging the Corrected Field Of Viewmentioning

confidence: 99%

Perspective: Wavefront shaping techniques for controlling multiple light scattering in biological tissues: Toward in vivo applications

Park

Lee

et al. 2018

APL Photonics

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Multiple light scattering has been regarded as a barrier in imaging through complex media such as biological tissues. Owing to recent advances in wavefront shaping techniques, optical imaging through intact biological tissues without invasive procedures can now be used for direct experimental studies, presenting promising application opportunities in in vivo imaging and diagnosis. Although most of the recent proof of principle breakthroughs have been achieved in the laboratory setting with specialties in physics and engineering, we anticipate that these technologies can be translated to biological laboratories and clinical settings, which will revolutionize how we diagnose and treat a disease. To provide insight into the physical principle that enables the control of multiple light scattering in biological tissues and how recently developed techniques can improve bioimaging through thick tissues, we summarize recent progress on wavefront shaping techniques for controlling multiple light scattering in biological tissues.

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“…Deep-tissue imaging faces the similar problem, and others have worked in this area of research showing that correction in conjugated-image planes is also possible for microscopy applications (33)(34)(35)(36).…”

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confidence: 99%

High-resolution in vivo imaging of mouse brain through the intact skull

Park

Sun

Cui

2015

Proc. Natl. Acad. Sci. U.S.A.

152

130

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Multiphoton microscopy is the current method of choice for in vivo deep-tissue imaging. The long laser wavelength suffers less scattering, and the 3D-confined excitation permits the use of scattered signal light. However, the imaging depth is still limited because of the complex refractive index distribution of biological tissue, which scrambles the incident light and destroys the optical focus needed for high resolution imaging. Here, we demonstrate a wavefrontshaping scheme that allows clear imaging through extremely turbid biological tissue, such as the skull, over an extended corrected field of view (FOV). The complex wavefront correction is obtained and directly conjugated to the turbid layer in a noninvasive manner. Using this technique, we demonstrate in vivo submicron-resolution imaging of neural dendrites and microglia dynamics through the intact skulls of adult mice. This is the first observation, to our knowledge, of dynamic morphological changes of microglia through the intact skull, allowing truly noninvasive studies of microglial immune activities free from external perturbations.neuroimaging | nonlinear microscopy | wavefront shaping | immunology | adaptive optics B reakthroughs in imaging technologies have been one of the major driving forces of new discoveries in biology (1). In the past two decades, we have witnessed that the advances of superresolution microscopy revolutionized our understanding of the complex dynamics in single cells (2). However, the techniques that work well on cultured cells often fail when being used to observe the cells in their native environment inside a living organism, the most ideal condition for biological studies. These difficulties result from the optical wavefront distortions induced by the inhomogeneous refractive index distribution in biological tissue.Because of this limitation, imaging inside deep tissue has been a challenging task, and the common goal of state of the art deep tissue-imaging methods is to recover the diffraction-limited resolution. These efforts can be categorized largely into two groups: the first capitalizes the use of longer wavelengths (3, 4) that undergo less scattering, whereas the second controls the optical wavefront, so that the wavefront distortions induced by the sample can be compensated (5-30). Longer wavelength excitation has demonstrated impressive penetration depths in the exposed brain (3) and imaging of the brain vascular structures beneath the skull (4). However, at the current state, using longer wavelengths is not a simple option because it requires special illumination sources [e.g., low repetition rate, high energy pulses (3)] or fluorescent probes (4), and provides only moderate spatial resolutions. In comparison, wavefront shaping allows the use of conventional light sources as well as the wide selection of wellestablished labels and functional indicators (31).Previous exciting works in adaptive optics (AO) have exploited these advantages and demonstrated aberration correction for a large field of view (FOV) by avera...

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Field of view advantage of conjugate adaptive optics in microscopy applications

Cited by 94 publications

References 28 publications

Widefield fluorescence microscopy with sensor-based conjugate adaptive optics using oblique back illumination

Widefield fluorescence microscopy with sensor-based conjugate adaptive optics using oblique back illumination

Perspective: Wavefront shaping techniques for controlling multiple light scattering in biological tissues: Toward in vivo applications

High-resolution in vivo imaging of mouse brain through the intact skull

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