Correcting spherical aberrations in a biospecimen using a transmissive liquid crystal device in two-photon excitation laser scanning microscopy

Tanabe, A.; Hibi, Terumasa; Ipponjima, Sari; Matsumoto, Kenji; Yokoyama, Masafumi; Kurihara, Makoto; Hashimoto, Nobuyuki; Nemoto, Tomomi

doi:10.1117/1.jbo.20.10.101204

Cited by 22 publications

(22 citation statements)

References 28 publications

(41 reference statements)

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“…The space-variant aberration can be restored by some deblurring algorithms [31], corrected by adaptive optics techniques [32,33], or reduced by use of objectives with a motorized correction collar. However, as we indicated above, to adjust RIs homogeneously is a straightforward and effective method to obtain deep tissue imaging with nanosheet wrapping a cleared tissue specimen.…”

Section: Resultsmentioning

confidence: 99%

Nanosheet wrapping-assisted coverslip-free imaging for looking deeper into a tissue at high resolution

et al. 2020

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In order to achieve deep tissue imaging, a number of optical clearing agents have been developed. However, in a conventional microscopy setup, an objective lens can only be moved until it is in contact with a coverslip, which restricts the maximum focusing depth into a cleared tissue specimen. Until now, it is still a fact that the working distance of a high magnification objective lens with a high numerical aperture is always about 100 μm. In this study, a polymer thin film (also called as nanosheet) composed of fluoropolymer with a thickness of 130 nm, less than one-thousandth that of a 170 μm thick coverslip, is employed to replace the coverslip. Owing to its excellent characteristics, such as high optical transparency, mechanical robustness, chemical resistance, and water retention ability, nanosheet is uniquely capable of providing a coverslip-free imaging. By wrapping the tissue specimen with a nanosheet, an extra distance of 170 μm for the movement of objective lens is obtained. Results show an equivalently high resolution imaging can be obtained if a homogenous refractive index between immersion liquid and mounting media is adjusted. This method will facilitate a variety of imaging tasks with off-the-shelf high magnification objectives.

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Section: Resultsmentioning

confidence: 99%

Nanosheet wrapping-assisted coverslip-free imaging for looking deeper into a tissue at high resolution

et al. 2020

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“…S3b and c), which corrected the spherical aberration. Moreover, to correct the remaining optical aberrations, it would be useful to adjust the RI of clearing reagents to 1.515 (Tanabe et al ., ; Ke et al ., ). Indeed, we previously proposed adaptive optics containing liquid crystal cells as an alternative approach to reducing various optical aberrations (Tanabe et al ., ).…”

Section: Discussionmentioning

confidence: 98%

Super‐resolution structural analysis of dendritic spines using three‐dimensional structured illumination microscopy in cleared mouse brain slices

Sawada

Kawakami

Shigemoto

et al. 2018

Eur J of Neuroscience

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Three‐dimensional (3D) super‐resolution microscopy technique structured illumination microscopy (SIM) imaging of dendritic spines along the dendrite has not been previously performed in fixed tissues, mainly due to deterioration of the stripe pattern of the excitation laser induced by light scattering and optical aberrations. To address this issue and solve these optical problems, we applied a novel clearing reagent, LUCID, to fixed brains. In SIM imaging, the penetration depth and the spatial resolution were improved in LUCID‐treated slices, and 160‐nm spatial resolution was obtained in a large portion of the imaging volume on a single apical dendrite. Furthermore, in a morphological analysis of spine heads of layer V pyramidal neurons (L5PNs) in the medial prefrontal cortex (mPFC) of chronic dexamethasone (Dex)‐treated mice, SIM imaging revealed an altered distribution of spine forms that could not be detected by high‐NA confocal imaging. Thus, super‐resolution SIM imaging represents a promising high‐throughput method for revealing spine morphologies in single dendrites.

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“…These devices, inserted between the objective lens and the revolver of a microscope, improved the quality of the fluorescent images in fixed-mouse-brain slices that were observed using two-photon excitation laser scanning microscopy, which were originally degraded by a spherical aberration. 4,5 However, asymmetric aberrations can also be induced by the shape of a biospecimen and a complicated refractive-index distribution in the sample, and degrade the optical performance considerably, even near the sample surface. [6][7][8] These asymmetric aberrations in biospecimens and their effects have been investigated experimentally by several authors.…”

Section: Introductionmentioning

confidence: 99%

Transmissive liquid-crystal device correcting primary coma aberration and astigmatism in laser scanning microscopy

et al. 2016

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Laser scanning microscopy allows 3D cross-sectional imaging inside biospecimens. However, certain aberrations produced can degrade the quality of the resulting images. We previously reported a transmissive liquid-crystal device that could compensate for the predominant spherical aberrations during the observations, particularly in deep regions of the samples. The device, inserted between the objective lens and the microscope revolver, improved the image quality of fixed-mouse-brain slices that were observed using two-photon excitation laser scanning microscopy, which was originally degraded by spherical aberration. In this study, we developed a transmissive device that corrects primary coma aberration and astigmatism, motivated by the fact that these asymmetric aberrations can also often considerably deteriorate image quality, even near the sample surface. The device's performance was evaluated by observing fluorescent beads using single-photon excitation laser scanning microscopy. The fluorescence intensity in the image of the bead under a cover slip tilted in the y-direction was increased by 1.5 times after correction by the device. Furthermore, the y-and z-widths of the imaged bead were reduced to 66% and 65%, respectively. On the other hand, for the imaged bead sucked into a glass capillary in the longitudinal x-direction, correction with the device increased the fluorescence intensity by 2.2 times compared to that of the aberrated image. In addition, the x-, y-, and z-widths of the bead image were reduced to 75%, 53%, and 40%, respectively. Our device successfully corrected several asymmetric aberrations to improve the fluorescent signal and spatial resolution, and might be useful for observing various biospecimens.

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Correcting spherical aberrations in a biospecimen using a transmissive liquid crystal device in two-photon excitation laser scanning microscopy

Cited by 22 publications

References 28 publications

Nanosheet wrapping-assisted coverslip-free imaging for looking deeper into a tissue at high resolution

Nanosheet wrapping-assisted coverslip-free imaging for looking deeper into a tissue at high resolution

Super‐resolution structural analysis of dendritic spines using three‐dimensional structured illumination microscopy in cleared mouse brain slices

Transmissive liquid-crystal device correcting primary coma aberration and astigmatism in laser scanning microscopy

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