Optical sectioning and high resolution visualization of trabecular meshwork using Bessel beam assisted light sheet fluorescence microscopy

Sandeep, C. S. Suchand; Sarangapani, Sreelatha; Hong, Xun; Aung, Tin; Baskaran, Mani; Murukeshan, Vadakke Matham

doi:10.1002/jbio.201900048

Cited by 11 publications

(11 citation statements)

References 41 publications

(44 reference statements)

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“…It is worth noting that, although utilized for single wavelength excitation, many of the LSFM systems found in the literature use semiapochromat (FLU) objectives [1, 6, 33–37]. Finally, because APO objectives are chromatically corrected for red, green, and blue colors, one may suppose that they are a good option for multicolor excitation LSFM [2, 12, 17, 23, 38–43] even when using Gaussian beams. However, the focus difference

(||, δ_{C^{'}} - δ_{F^{'}})

ranges from 2.29 to 14.29 μm, when using low NAs from 0.4 to 0.16.…”

Section: Resultsmentioning

confidence: 99%

Multicolor light‐sheet microscopy for a large field of view imaging: A comparative study between Bessel and Gaussian light‐sheets configurations

Luna‐Palacios

Licea-Rodríguez

Camacho-Lopez

et al. 2022

Journal of Biophotonics

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Light‐sheet fluorescence microscopy (LSFM) is useful for developmental biology studies, which require a simultaneous visualization of dynamic microstructures over large fields of views (FOVs). A comparative study between multicolor Bessel and Gaussian‐based LSFM systems is presented. Discussing the chromatic implications to achieve colocalized and large FOVs when both optical arrays are implemented under the same excitation objective is the purpose of this work. The light‐sheets FOVs, optical sectioning, and resolution are experimentally characterized and discussed. The advantages of using Bessel beams and the main drawbacks of using Gaussian beams for multicolor imaging are highlighted. Multiple Bessel excitation minimizes the FOV's mismatch's effects due to the beams chromatic defocusing and alleviates the aside object blurring obtained with multiple Gaussian beams. It also offers a fair homogeneous axial resolution and optical sectioning over a larger effective FOV. Imaging over perithecia samples of the fungus Sordaria macrospora demonstrates such advantages. This work complements previous comparative studies that discuss only single wavelengths light‐sheets excitations.

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(||, δ_{C^{'}} - δ_{F^{'}})

ranges from 2.29 to 14.29 μm, when using low NAs from 0.4 to 0.16.…”

Section: Resultsmentioning

confidence: 99%

Multicolor light‐sheet microscopy for a large field of view imaging: A comparative study between Bessel and Gaussian light‐sheets configurations

Luna‐Palacios

Licea-Rodríguez

Camacho-Lopez

et al. 2022

Journal of Biophotonics

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“…By sweeping the Bessel beam in x-direction to project a light sheet at each z -plane, a 3D volumetric stack can be created. 51 The sample under study should be kept stationary throughout this 3D data acquisition, which will help in minimizing motion artifacts. Such motion artefacts will be lesser in human eye imaging compared with animals under anesthesia.…”

Section: Discussionmentioning

confidence: 99%

Noninvasive and Noncontact Sequential Imaging of the Iridocorneal Angle and the Cornea of the Eye

Hong

Sandeep

Shinoj

et al. 2020

Trans. Vis. Sci. Tech.

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High-resolution imaging of the critical anatomic structures of the eye, especially of the anterior chamber, in vivo, remains a challenge, even with currently available state-of-the-art medical imaging techniques. This study aims for the noninvasive and noncontact sequential imaging of the iridocorneal angle, especially the trabecular meshwork (TM) and the cornea of the eye in high-resolution using a newly developed imaging platform. Methods: Bessel beam scanned light sheet fluorescence microscopy is used to attain high-resolution images of the TM. The ability of the Bessel beam to self-reconstruct around obstacles increases the image contrast at the TM region inside eye by reducing scattering and shadow artifacts. With minimal modifications, the excitation arm of the developed imaging system is adapted for noncontact, high-resolution corneal imaging. Results: High-resolution images of the TM structures and cellular-level corneal structures are obtained in ex vivo porcine eyes, and subsequently in New Zealand white rabbit, in vivo. The spatial resolution of the developed system is 2.19 μm and has a noncontact working distance of 20 mm. Conclusions: A high-resolution imaging platform for noncontact sequential imaging of the TM and the cornea of the eye is developed. This imaging system is expected to be of potential interest in the evaluation and diagnosis of glaucoma and corneal diseases. Translational Relevance: The developed prototype offers the plausibility of in vivo, noncontact, and high-resolution imaging of the iridocorneal angle and cornea of the eye that will aid clinicians in diagnosing open-angle glaucoma and corneal diseases better.

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“…The Bessel beam's self-healing capability, even in a scattering environment, aids in maintaining relatively uniform illumination, resulting in improved image contrast. to the one reported in reference [59], except for the excitation objective. The use of a longerworking-distance (30 mm) apochromatic objective for exciting fluorescence from the TM provided much more flexibility during the measurements.…”

Section: Bessel Beam-based Light Sheet Fluorescence Microscopy (Lsfm)...mentioning

confidence: 91%

“…Figure 5 shows the reported sequen aging system utilizing Bessel beam-based LSFM for TM imaging [72]. Suchand Sandeep et al reported the three-dimensional (3D) visualization of the TM on an intact ex vivo porcine eye utilizing a modified version of the Bessel beam-based system [59]. The system developed had a lateral resolution of 0.78 μm and an axial resolution of 2.2 μm.…”

Section: Bessel Beam-based Light Sheet Fluorescence Microscopy (Lsfm)...mentioning

confidence: 99%

Bessel Beams in Ophthalmology: A Review

Suchand Sandeep,

Khairyanto,

Aung

et al. 2023

Micromachines

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The achievable resolution of a conventional imaging system is inevitably limited due to diffraction. Dealing with precise imaging in scattering media, such as in the case of biomedical imaging, is even more difficult owing to the weak signal-to-noise ratios. Recent developments in non-diffractive beams such as Bessel beams, Airy beams, vortex beams, and Mathieu beams have paved the way to tackle some of these challenges. This review specifically focuses on non-diffractive Bessel beams for ophthalmological applications. The theoretical foundation of the non-diffractive Bessel beam is discussed first followed by a review of various ophthalmological applications utilizing Bessel beams. The advantages and disadvantages of these techniques in comparison to those of existing state-of-the-art ophthalmological systems are discussed. The review concludes with an overview of the current developments and the future perspectives of non-diffractive beams in ophthalmology.

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Optical sectioning and high resolution visualization of trabecular meshwork using Bessel beam assisted light sheet fluorescence microscopy

Cited by 11 publications

References 41 publications

Multicolor light‐sheet microscopy for a large field of view imaging: A comparative study between Bessel and Gaussian light‐sheets configurations

Multicolor light‐sheet microscopy for a large field of view imaging: A comparative study between Bessel and Gaussian light‐sheets configurations

Noninvasive and Noncontact Sequential Imaging of the Iridocorneal Angle and the Cornea of the Eye

Bessel Beams in Ophthalmology: A Review

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