A planar Airy beam light-sheet for two-photon microscopy
Neveen A. Hosny, James A. Seyforth, Gunnar Spickermann, Thomas J. Mitchell, Pedro Almada, Robert Chesters, Scott J. Mitchell, George Chennell, Anthony C. Vernon… 展开作者
Neveen A. Hosny, James A. Seyforth, Gunnar Spickermann, Thomas J. Mitchell, Pedro Almada, Robert Chesters, Scott J. Mitchell, George Chennell, Anthony C. Vernon, Kwangwook Cho, Deepak P. Srivastava, Robert Forster, Tom Vettenburg
We demonstrate the first planar Airy light-sheet microscope. Fluorescence light-sheet microscopy has become the method of choice to study large biological samples with cellular or sub-cellular resolution. The propagation-invariant Airy beam enables a ten-fold increase in field-of-view with single-photon excitation; however, the characteristic asymmetry of the light-sheet limits its potential for multi-photon excitation. Here we show how a planar light-sheet can be formed from the curved propagation-invariant Airy beam. The resulting symmetric light sheet excites two-photon fluorescence uniformly across an extended field-of-view without the need for deconvolution. We demonstrate the method for rapid two-photon imaging of large volumes of neuronal tissue.
We build a simulation platform for Airy beam photoacoustic microscopy based on K-Wave simulation toolbox in MATLAB. The sample was located in the middle of z direction in the environment. The propagation of Airy beam irradiated on the sample was simulated by solving the Schrodinger equation. And the intensity of Airy beam was simulated by Airy wave pocket. The simulation result showed that: in A-scan, the intensity of light waved over time, reached peak at 49ns; in C-scan, the image agrees with the sample. This research showed that the Airy beam with large depth of field and no diffraction can…
Super-resolution imaging techniques enable nanoscale microscopy in fields such as physics, biology, and chemistry. However, many super-resolution techniques require specialized optical components, such as a helical-phase mask. We present a novel technique, Super-resolution Airy disk Microscopy (SAM) that can be used in a standard confocal microscope without any specialized optics. We demonstrate this technique, in combination with ground state depletion, to image and control nitrogen-vacancy (NV) centers in bulk diamond below the diffraction limit. A greater than 14-fold improvement in resolut…
Scattering and absorption limit the penetration of optical fields into tissue, but wavefront correction, often used to compensate for these effects, is incompatible with wide field-of-view imaging and complex to implement. We demonstrate a new approach for increased penetration in light-sheet imaging, namely attenuation-compensation of the light field. This tailors an exponential intensity increase along the illuminating propagation-invariant field, enabling the redistribution of intensity strategically within a sample. This powerful yet straightforward concept, combined with the self-healing …
Sample-induced image-degradation remains an intricate wave-optical problem in light-sheet microscopy. Here we present biobeam, an open-source software package that enables to simulate operational light-sheet microscopes by combining data from 10^5 - 10^6 multiplexed and GPU-accelerated point-spread-function calculations. The wave-optical nature of these simulations leads to the faithful reproduction of spatially varying aberrations, diffraction artifacts, geometric image distortions, adaptive optics, and emergent wave optical phenomena, and renders image-formation in light-sheet microscopy com…
We demonstrate curved modifications with lengths of up to 2 mm within borosilicate glass produced by single 1030 nm picosecond laser shots with an Airy beam profile. Plasma ignition in the side lobes of the beam as well as surface damage prove to be the crucial limitations for confined bulk energy deposition on a curved trajectory. A combined experimental and numerical analysis reveals optimum laser parameters for confined bulk energy deposition. This way we achieved single pass cutting of a 525 $μ$m thick glass sheet with a well defined convex edge down to a bending radius of 774 $μ$m.
High resolution light-sheet optical microscopy requires illuminating a sample by a thin excitation light sheet with large area. Here, we describe methods for producing light sheets with extended length, as compared to Gaussian-beam-type diffracting light sheets with the same thickness, without scanning or dithering the light beams.
Widefield Nanodiamond Quantum Sensing Based on Light-Sheet Microscopy
The Chinese University of Hong Kong（香港中文大学）；Hefei University of Technology（合肥工业大学）；Centre for Quantum Coherence, The Chinese University of Hong Kong（香港中文大学量子相干研究中心）；The State Key Laboratory of Quantum Information Technologies and Materials, The Chinese University of Hong Kong（香港中文大学量子信息技术与材料国家重点实验室）；New Cornerstone Science Laboratory, The Chinese University of Hong Kong（香港中文大学新基石科学实验室）
An asymmetric \b{eta}-sheet structure of spider silk is said to induce optical second harmonic generation. In this paper, using an in-house non-scanning type femtosecond pulse laser second harmonic generation microscope, we characterized the behavior of the \b{eta}-sheet of spider silk under an applied external force. The orientation of the \b{eta}-sheets was more unidirectional when the silk was extended. One of the origins of the high mechanical strength of the dragline is suggested to be the physical arrangement of its \b{eta}-sheets.
Oblique plane microscopy (OPM) is a single objective light-sheet microscopy which performs three dimensional (3D) imaging by axial scan of the generated light-sheet. Recently, multiple techniques for lateral scan of the generated light-sheet in OPM have emerged. However, their suitability for geometrically distortion free 3D imaging, which essentially requires a constant tilt light-sheet scan, has not been evaluated. In this work, we use a geometrical optics approach and derive analytical relationship for the amount of tilt variance in planar mirror based scanned oblique plane illumination (SO…
Asymmetric $\rm β$-sheet protein structures in spider silk should induce nonlinear optical interaction such as second harmonic generation (SHG) which is experimentally observed for a radial line and dragline spider silk by using an imaging femtosecond laser SHG microscope. By comparing different spider silks, we found that the SHG signal correlates with the existence of the protein $\rm β$-sheets. Measurements of the polarization dependence of SHG from the dragline indicated that the $\rm β$-sheet has a nonlinear response depending on the direction of the incident electric field. We propose a …
This Letter presents the first demonstration of multi-tile stitching for large scale 3D imaging in single objective light-sheet microscopy. We show undistorted 3D imaging spanning complete zebrafish larvae, and over 1 mm3 volumes for thick mouse brain sections. We use remote galvo scanning for light-sheet creation and develop a processing pipeline for 3D tiling across different axes. With the improved one photon (1p) tilt-invariant scanned oblique plane illumination (SOPi) microscope presented here, we demonstrate cellular resolution imaging at depths exceeding 330 um in optically scattering m…
We report on a very first experimental observation of a new curved plasmonic beam, a plasmonic hook (PH), for surface plasmon-polariton (SPP) waves. We demonstrate that the SPP PH effect could be realized within a trapezoidal dielectric microstructure with the relative ease of fabrication compared to the generation of the Airy SPP waves which require complex techniques to compensate the wave vectors mismatch. Using numerical analysis, we predicted the PH phenomenon, optimized the microstructure and then confirmed the SPP PH existence via amplitude and phase-resolved scattering scanning near-fi…
We report second- and third-harmonic generation in monolayer MoS$_\mathrm{2}$ as a tool for imaging and accurately characterizing the material's nonlinear optical properties under 1560 nm excitation. Using a surface nonlinear optics treatment, we derive expressions relating experimental measurements to second- and third-order nonlinear sheet susceptibility magnitudes, obtaining values of $|χ_s^{(2)}|=2.0\times10^{-20}$ m$^2$ V$^{-1}$ and for the first time for monolayer MoS$_\mathrm{2}$, $|χ_s^{(3)}|=1.7\times10^{-28}$ m$^3$ V$^{-2}$. These sheet susceptibilities correspond to effective bulk n…
Light-sheet microscopy (LSM) is a powerful imaging technique that uses a planar illumination oriented orthogonally to the detection axis. Two-photon (2P) LSM is a variant of LSM that exploits the 2P absorption effect for sample excitation. The light polarization state plays a significant, and often overlooked, role in 2P absorption processes. The scope of this work is to test whether using different polarization states for excitation light can affect the detected signal levels in 2P LSM imaging of typical biological samples with a spatially unordered dye population. Supported by a theoretical …
Fluorescent imaging plays a critical role in a myriad of scientific endeavors, particularly in the biological sciences. Three-dimensional imaging of fluorescent intensity often requires serial data acquisition, that is voxel-by-voxel collection of fluorescent light emitted throughout the specimen with a non-imaging single-element detector. While non-imaging fluorescence detection offers some measure of scattering robustness, the rate at which dynamic specimens can be imaged is severely limited. Other fluorescent imaging techniques utilize imaging detection to enhance collection rates. A notabl…
Two-photon light-sheet live imaging at kilohertz frame rate using birefringence-based pulse splitting
Laboratoire d’Optique et Biosciences, CNRS, INSERM, École Polytechnique, Institut Polytechnique de Paris（光学与生物科学实验室，法国国家科学研究中心，法国国家健康与医学研究院，巴黎综合理工学院，巴黎理工学院）；ICube Research Institute, CNRS, University of Strasbourg（ICube研究所，法国国家科学研究中心，斯特拉斯堡大学）；Sorbonne Université, CNRS, Laboratoire Jean Perrin（索邦大学，法国国家科学研究中心，让·佩兰实验室）
Structured illumination microscopy (SIM) provides images of fluorescent objects at an enhanced resolution greater than that of conventional epifluorescence wide-field microscopy. Initially demonstrated in 1999 to enhance the lateral resolution two-fold, it has since been extended to enhance axial resolution two-fold (2008), applied to live-cell imaging (2009) and combined with myriad other techniques, including interferometric detection (2008), confocal microscopy (2010) and light sheet illumination (2012). Despite these impressive developments, SIM remains, perhaps, the most poorly understood…
Light sheet fluorescence microscopy provides optical sectioning and is widely used in volumetric imaging of large specimens. However, the axial resolution and the lateral Field of View (FoV) of the system, defined by the light sheet, typically limit each other due to the spatial band product of the excitation objective. Here, we develop a simple multi-focus scheme to extend the FoV, where a Gaussian light sheet can be focused at three or more consecutive positions. Axially overlapped multiple light sheets significantly enlarge the FoV with improved uniformity and negligible loss in axial resol…
Structured light has revolutionized optical particle manipulation and nano-scale material processing. In particular, propagation-invariant structured light fields, such as Bessel beams, have enabled applications that require robust intensity distributions. Their self-healing nature facilitates imaging with enhanced resolution e.g. in light-sheet microscopy. The prominent high-intensity features of propagation-invariant fields such as Airy, Bessel, and Mathieu beams can be understood in terms of caustics. While these beams have found many applications in material processing and trapping, these …
Department of Electrical Engineering, University of Notre Dame（圣母大学电气工程系）；Department of Biological Sciences, University of Notre Dame（圣母大学生物科学系）
Fluorescence microscopy has been a significant tool to observe long-term imaging of embryos (in vivo) growth over time. However, cumulative exposure is phototoxic to such sensitive live samples. While techniques like light-sheet fluorescence microscopy (LSFM) allow for reduced exposure, it is not well suited for deep imaging models. Other computational techniques are computationally expensive and often lack restoration quality. To address this challenge, one can use various low-dosage imaging techniques that are developed to achieve the 3D volume reconstruction using a few slices in the axial …
Amyloid proteins are associated with a broad spectrum of neurodegenerative diseases. However, it remains a grand challenge to extract molecular structure information from intracellular amyloid proteins in their native cellular environment. To address this challenge, we developed a computational chemical microscope integrating 3D mid-infrared photothermal imaging with fluorescence imaging, termed Fluorescence-guided Bond-Selective Intensity Diffraction Tomography (FBS-IDT). Based on a low-cost and simple optical design, FBS-IDT enables chemical-specific volumetric imaging and 3D site-specific m…
The aberrations induced by the sample and/or by the sample holder limit the resolution of optical microscopes. Wavefront correction can be achieved using a deformable mirror with wavefront sensorless optimization algorithms but, despite the complexity of these systems, the level of correction is often limited to a small area in the field of view of the microscope. In this work, we present a plug and play module for aberration measurement and correction. The wavefront correction is performed through direct wavefront reconstruction using the spinning-pupil aberration measurement and controlling …
In conventional Kerr- and Faraday microscopy the sample is illuminated with plane-polarised light and a magnetic domain contrast is generated by an analyser making use of the Kerr- or Faraday rotation. In this paper we demonstrate possibilities of analyser-free magneto-optical microscopy based on magnetisation-dependent intensity modulations of the light: (i) The transverse Kerr effect can be applied for in-plane magnetised material, demonstrated for an FeSi sheet. (ii) Illuminating the same sample with circularly polarised light leads to a domain contrast with a different symmetry as the conv…
