Stochastic optical reconstruction microscopy (STORM) achieves super-resolution imaging by blinking individual dye molecules in thiol-containing media. While STORM is well-established for imaging thin biological specimens, its application to thick tissues has been limited by light scattering. Mounting media with an oil refractive index have shown promise in improving imaging depth and resolution in optical microscopy, but such buffers have not been explored for STORM. Here, we present a 3-pyridinemethanol-based STORM buffer with a refractive index matching standard immersion oil. Our buffer demonstrates comparable performance to conventional STORM buffers and exceptional stability for 5 weeks, enabling imaging of numerous cells on a single slide and larger field-of-view imaging. With perfect index matching, our buffer simplifies imaging and holds potential for lightsheet STORM applications in thick tissues.
Phase-only spatial light modulators (SLMs) are extensively utilized for controlling the phase of light in diverse applications. However, liquid-crystal-on-silicon (LCOS) SLMs exhibit undesired spatial variations in both phase response and optical flatness across the SLM panel, which necessitates calibration for achieving accurate phase control. Here, we propose a rapid and straightforward calibration approach to address these non-uniformities at the single-pixel level. Our method leverages Twyman-Green interferometry without relying on a piezoelectric transducer, significantly reducing the measurement time to approximately 4.5 seconds by collecting only 18 interferograms of SLM patterns with constant gray levels. This remarkable speed ensures minimal vulnerability to environmental disturbances during interferometry. We provide a comprehensive description of the calibration procedure, evaluate the performance of the calibrated local pixel response, and validate the flatness calibration using SLM-based phase shift interferometry.
Oblique plane microscopy-based single molecule localization microscopy (obSTORM) shows promise for superresolution imaging in thick biological samples. However, the Gaussian point spread function (PSF) model used in previous studies limits imaging resolution and axial localization range in obSTORM due to poor fitting with actual PSF shapes. To overcome these limitations, we employed cubic splines to construct a more precise PSF model. This refined model enhances three-dimensional (3D) localization precision, improving obSTORM imaging of mouse retina tissues. It increases imaging resolution by approximately 1.2 times, enables seamless stitching of single molecules across optical sections, and doubles the sectional interval in volumetric obSTORM imaging by extending the usable section thickness. The cubic spline PSF model offers a promising approach for achieving faster and more accurate volumetric obSTORM imaging of biological specimens.
Asymmetric transport is an uneven physical response of counter-propagating signals that has significantly contributed to fundamental science and revolutionized advanced technology via a variety of significant devices including diodes and isolators in electronics, optics, acoustics, and heat transfer. Photonic metasurfaces are two-dimensional ultrathin arrays of engineered subwavelength meta-atoms, acting as local phase shifters, which unprecedentedly mold wavefronts at will with a virtually flat optical element. While such an architecture can be potentially harnessed to achieve two-way asymmetric response of free-space light at an optically thin flatland, asymmetric light transport cannot be fundamentally achieved by any linear system including linear metasurfaces. Here, we report asymmetric transport of free-space light at nonlinear metasurfaces, with harmonic generation, upon transmission and reflection. We also derive the nonlinear generalized Snell’s laws of reflection and refraction which were experimentally verified by angle-resolved anomalous refraction and reflection of the nonlinear light. The asymmetric transport at optically thin nonlinear interfaces is revealed by comparing the original path of light through the metasurface with its corresponding reversed propagation path. Such a two-way asymmetric response at metasurfaces opens a new paradigm for free-space ultrathin lightweight optical devices with one-way operation including unrivaled optical valves and diodes.
While single-molecule localization microscopy (SMLM) offers superior super-resolution for biology, typical SMLM system using highly-inclined off-axis illumination limits an imaging depth to only a few microns from a coverslip surface. Alternative SMLM system using light-sheet illumination has extended the accessible depth for whole cells or small embryos, but may be less practical as it requires specialized or dedicated sample devices. Furthermore, for typical tissue samples (laterally a few millimeters or wider), the lateral lightsheet illumination is no longer applicable. Here, we demonstrate oblique light-sheet SMLM (obSTORM) that provides a facile and practical platform with a full compatibility with tissue samples. By using a single-objective, inclined lightsheet and directly detecting single molecules along the oblique plane, obSTORM opens new doors for tissue-level super-resolution imaging.
The formation of half-light half-matter quasiparticle exciton polariton and its condensation in semiconductor microcavities are striking phenomena for the macroscopically quantum coherence at elevated temperature. The matter constituent of exciton polariton dictates the interacting behaviors of these bosonic particles primarily via exciton-exciton interactions. However, these interactions are all limited to the ground state exciton, although they are expected to be much stronger at Rydberg states with higher principal numbers. Here, for the first time, we observe the spontaneous formed Rydberg exciton polaritons (REPs) in a high quality Fabry-Perot cavity embedded with single crystal inorganic perovskite. Such REPs exhibit strong nonlinear behavior and anisotropic, enabling an anomalous dynamic process that leads to a coherent polariton condensate with prominent blue shift. This discovery presents a unique platform to study quantum coherent many-body physics, and enables unprecedented manipulation of these Rydberg states by new means such as chemical composition engineering, structural phase control, and external gauge fields. The solid state REP and its condensates also hold great potential for important applications, such as sensing, communication, and quantum computing.
KEYWORDS: Point spread functions, Optical transfer functions, Imaging systems, 3D image processing, Objectives, Mirrors, Diffraction, Spectral resolution, Microscopy, 3D acquisition
Direct oblique plane imaging is a high-speed microscopy technique that observes a sample’s plane that is inclined to the focal plane of the microscope objective lens. This wide-field microscopy is suitable for a study of fast dynamics of living samples where the principle plane of interest is tilted to the focal plane. A way to implement this imaging technique is to use remote focusing together with a tilted mirror, which involves asymmetrical pupil function of the imaging system. We rigorously study the anisotropic resolving power of the oblique plane imaging using a vectorial diffraction theory. From the derived effective pupil function, we calculate vectorial point spread function (PSF) and optical transfer function (OTF). We show that the two-dimensional (2D) PSF of the direct oblique plane imaging is not merely an oblique crosssection of the 3D PSF of circular aperture system. Similarly, 2D OTF of the oblique plane imaging is different from 2D oblique projection of conventional 3D OTF in circular aperture system.
New confocal microscopy having no mechanical beam scanning devices is proposed. The proposed system can get two-dimensional information of a specimen in real-time by using spectral encoding technique and slit aperture. Spectral encoding technique is used to encode one- dimensional lateral information of the specimen in wavelength by a diffraction grating and a broadband light source. The modeling of the optical system is conducted. The effect of slit width variation on the axial response of the system is evaluated by numerical simulation based on the wave optics. Proper width of the slit aperture which plays a crucial role of the out-of-focus blur rejection is determined by a compromise between axial resolution and signal intensity from the simulation result. Design variables and governing equations of the system are derived on the assumption of a lateral sampling resolution of 50 nm. The system is designed to have a mapping error less than the half pixel size, to be diffraction-limited and to have the maximum illumination efficiency. The designed system has a FOV of 12.8 μm x 9.6 μm, a theoretical axial FWHM of 1.1 μm and a lateral magnification of -367.8.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.