High Numerical Aperture Objective Lens Research Articles

Enhancing Bioluminescence Imaging of Cultured Tissue Explants Using Optical Telecompression.

Long-term observation of single-cell oscillations within tissue networks is now possible by combining bioluminescence reporters with stable tissue explant culture techniques. This method is particularly effective in revealing the network dynamics in systems with slow oscillations, such as circadian clocks. However, the low intensity of luciferase-based bioluminescence requires signal amplification using specialized cameras (e.g., I-CCDs and EM-CCDs) and prolonged exposure times, increasing baseline noise and reducing temporal resolution. To address this limitation, we implemented a cost-effective optical enhancement technique called telecompression, first used in astrophotography and now commonly used in digital photography. By combining a high numerical aperture objective lens with a magnification-reducing relay lens, we significantly increased the collection efficiency of the bioluminescence signal without raising the baseline CCD noise. This method allows for shorter exposure times in time-lapse imaging, enhancing temporal resolution and enabling more precise period estimations. Our implementation demonstrates the feasibility of telecompression for enhancing bioluminescence imaging for the tissue-level network observation of circadian clocks.

Generation of variable light fields by radially polarized chirped circular Airy vortex beams

Based on Richards-Wolf vector diffraction theory, the tight focusing properties of radially polarized chirped circular Airy vortex beams (RP-CCAVBs) are theoretically investigated using a high numerical aperture objective lens in the paper. The results shown that super-resolution scalable optical needles, dark channels, elliptical optical cages, and optical oscillations can be obtained by adjusting the scale factor, exponential attenuation coefficient, main ring radius, and topological charge of optical vortex in the incident RP-CCAVBs. The DOF of optical needle and dark channels is exponent dependent on scaling factor ω of RP-CCAVBs. And the position optical needle and dark channels can be moved by changing chirped parameter of RP-CCAVBs. The size of obtained optical cages can be adjusted by changing the radius of main rings in RP-CCAVBs. The intensity distribution of the obtained optical oscillations is intensely affected by parameter a and chirp parameter of RP-CCAVBs.

Creation and manipulation of optical Meron topologies in tightly focused electromagnetic field

The optical topology, which serves as a stable spatial electromagnetic structure, offers a new dimension for applications in the field of optical information processing, transmission, and storage. In recent years, there has been an increasing focus on these spatially structured light fields. By reversing the radiation of orthogonal dipole pairs, we propose an approach to generate Meron topologies within the focused light field while also investigating the evolution of the Meron structure along the longitudinal axis. Through introducing a dipole placed along the z-axis, we achieve precise positioning and fine adjustment of the topological center. The stability of Meron under a high numerical aperture objective lens (NA = 0.95) can be effectively demonstrated.

Astigmatism-based active focus stabilisation with universal objective lens compatibility, extended operating range and nanometer precision

Focus stabilisation is vital for long-term fluorescence imaging, particularly in the case of high-resolution imaging techniques. Current stabilisation solutions either rely on fiducial markers that can be perturbative, or on beam reflection monitoring that is limited to high-numerical aperture objective lenses, making multimodal and large-scale imaging challenging. We introduce a beam-based method that relies on astigmatism, which offers advantages in terms of precision and the range over which focus stabilisation is effective. This approach is shown to be compatible with a wide range of objective lenses (10x-100x), typically achieving <10 nm precision with >10 μm operating range. Notably, our technique is largely unaffected by pointing stability errors, which in combination with implementation through a standalone Raspberry Pi architecture, offers a versatile focus stabilisation unit that can be added onto most existing microscope setups.

Fabrication of microneedles using two photon-polymerization with low numerical aperture

Three-dimensional (3D) micro-nano fabrication through two-photon polymerization (TPP) is a prominent approach for fabricating complex 3D structures. However, the existing techniques face limitations, such as small fabrication area, dependence on high numerical aperture (e.g., oil immersion) objective lenses, which are more expensive. We developed a 780 nm fs laser system integrated with a 2D galvanometer system and Z axis to create a 3D micro-nano printing system. A method to print arbitrarily complex 3D structures while maintaining an extremely high lateral spatial resolution of sub-100nm is proposed by using a cheap low numerical aperture objective lens with the laser power and exposure time precisely controlled. The impact of the laser power and exposure time on the resolution of 3D micro-nano processing is investigated, and we identify the optimal processing environment for achieving the best resolution. The printed linewidth of 62 nm was achieved without the use of an oil immersed objective lens, and only one laser beam was used instead of stimulated emission depletion. Microneedle arrays for animal injection were fabricated using this strategy, which can improve the performance of animal organoid repair. Our results suggest that TPP micro-nano fabrication is an effective method for processing microneedle arrays for biomedical engineering, with the ability to improve the processing resolution with a cheap low numerical aperture objectives lens (e.g., NA = 0.4) through meticulous control of laser power and exposure time. This approach opens up possibilities for affordable and scalable fabrication in sub-100nm structures.

Angle-resolving spectral ellipsometry using structured light for direct measurement ofellipsometric parameters.

We propose a compact angle-resolving spectral ellipsometry. Using the structured light generated from a digital micro-mirror device (DMD), what we believe to be a novel pattern is illuminated to the back focal plane of the high numerical aperture (NA) objective lens. As a result, ellipsometric parameters with fine resolution of both the wavelength and incidence angle domain can be directly measured. The incidence angle can be resolved by resolution under 1° ranging from 35° to 59° by the radius of the projected images. A spectrometer as a detector enables acquisition by the resolution of 0.7nm from 410 to 700nm, and the fiber reduces measurement spot size to a single micrometer. Additionally, the measurement process does not require any rotating optical components or moving parts, needing only digital modification of the projected image. This simplifies the sequences and reduces the measurement time. The 2D (angle of incidence and spectral domain) ellipsometric parameter plane measured by the proposed method was used to measure the thickness of various samples. The measurement result was verified in comparison with a commercial ellipsometer. The accuracy and precision of the result show that the proposed method is capable of precise measurement of thin films.

Evolution and particle trapping dynamics of circular Pearcey-Airy Gaussian vortex beams in tightly focused systems.

This study investigates the propagation and evolution of self-focusing circular Pearcey-Airy Gaussian vortex beams (CPAGVB) through high numerical aperture objective lenses. CPAGVB demonstrates a unique light field distribution compared to the circular Pearcey vortex beam and circular Airy Gaussian vortex beam. By adjusting optical distribution factors, main radii, and off-axis vortex pair positions, a variety of light field structures can be generated, including asymmetric micro-optical bottles, quasi-flat-top beam micro-optical bottles, and dual optical bottles. The particle trapping performance of CPAGVB is examined, revealing a gradient force eight orders of magnitude larger than its scattering force, up to twice the peak gradient force, and 2.5 times the scattering force of CAGVB. Further analysis of lateral power flow density, spin density vector, and total angular momentum distribution at the focal plane unveils the dynamics of particle motion toward the center. The Gouy phase difference under varying main radii reveals two types of normalized spin density vectors, characterized by helical and oscillating distributions. Additionally, the study examines the two-dimensional polarization ellipse distribution at the focal plane, elucidating the formation of central polarization singularities with axial vortices and the impact of peripheral polarization rearrangement on phase singularities. This research advances the comprehension of CPAGVB's distinctive properties and potential applications in micro-optical systems and particle manipulation.

Mesoscale standing wave imaging.

Standing wave (SW) microscopy is a method that uses an interference pattern to excite fluorescence from labelled cellular structures and produces high-resolution images of three-dimensional objects in a two-dimensional dataset. SW microscopy is performed with high-magnification, high-numerical aperture objective lenses, and while this results in high-resolution images, the field of view is very small. Here we report upscaling of this interference imaging method from the microscale to the mesoscale using the Mesolens, which has the unusual combination of a low-magnification and high-numerical aperture. With this method, we produce SW images within a field of view of 4.4mm × 3.0mm that can readily accommodate over 16,000 cells in a single dataset. We demonstrate the method using both single-wavelength excitation and the multi-wavelength SW method TartanSW. We show application of the method for imaging of fixed and living cells specimens, with the first application of SW imaging to study cells under flow conditions.

In-situ calibration of the objective lens of an angle-resolved scatterometer for nanostructure metrology.

Due to the advantages of being non-contact, non-destructive, highly efficient, and low in cost, scatterometry has emerged as a powerful technique for nanostructure metrology. In this paper, we propose an angle-resolved scatterometer composed of a scattered light acquisition channel and a spatial imaging channel, which is capable of detecting multi-order diffracted light in a single measurement. Since the high numerical aperture objective lens is usually employed in an angle-resolved scatterometer, the polarization effect of the objective lens introduced by the non-normal incidence and installation stress should be considered. An in-situ calibration method for the objective lens's polarization effects is proposed, in which a known analyzer is appended to the output light path to enable the extraction of the ellipsometric parameters of isotropic samples. Then the polarization effect of the objective lens can be determined in-situ by fitting the measured ellipsometric parameters to the calculated ones. With the objective lens polarization effect being considered, significant improvements in the accuracy and repeatability precision can be achieved in the metrology of the film thickness and grating topography parameters.

High-throughput single-molecule localization microscopy: Potential clinical applications.

High-throughput single-molecule localization microscopy: Potential clinical applications.

Tight Focus Vector Light Field of Hybrid Double Circularly Polarized Beams

Based on the Richard-Wolf diffraction theory, we demonstrate the tightly focused optical field distribution with two beams circularly polarized light. We deduced the expression of the double-beam superimposed vortex light intensity of left-handed circularly polarized light and right-handed circularly polarized light under the focusing condition of a high numerical aperture objective lens. Based on the different vortex topological charge values, amplitudes and initial phases of the dual beams, we numerically simulated the light intensity distribution of the superimposed hybrid focused light field. It is found that a variety of peculiar vortex spots can be obtained through the tight focus stacking of the double beams. These complex hybrid focused light fields with different polarizations can be used in fields such as optical field micro-manipulation or nonlinear fluorescence microscopy imaging.

Improving two-photon excitation microscopy for sharper and faster biological imaging.

Two-photon excitation laser scanning microscopy (TPLSM) has provided many insights into the life sciences, especially for thick biological specimens, because of its superior penetration depth and less invasiveness owing to the near-infrared wavelength of its excitation laser light. This paper introduces our four kinds of studies to improve TPLSM by utilizing several optical technologies as follows: (1) A high numerical aperture objective lens significantly deteriorates the focal spot size in deeper regions of specimens. Thus, approaches to adaptive optics were proposed to compensate for optical aberrations for deeper and sharper intravital brain imaging. (2) TPLSM spatial resolution has been improved by applying super-resolution microscopic techniques. We also developed a compact stimulated emission depletion (STED) TPLSM that utilizes electrically controllable components, transmissive liquid crystal devices, and laser diode-based light sources. The spatial resolution of the developed system was five times higher than conventional TPLSM. (3) Most TPLSM systems adopt moving mirrors for single-point laser beam scanning, resulting in the temporal resolution caused by the limited physical speed of these mirrors. For high-speed TPLSM imaging, a confocal spinning-disk scanner and newly-developed high-peak-power laser light sources enabled approximately 200 foci scanning. (4) Several researchers have proposed various volumetric imaging technologies. However, most technologies require large-scale and complicated optical setups based on deep expertise for microscopic technologies, resulting in a high threshold for biologists. Recently, an easy-to-use light-needle-creating device was proposed for conventional TPLSM systems to achieve one-touch volumetric imaging.

Development of pupil divided optics with magnification correction for high NA oblique imaging

For manufacturing semiconductor devices and hard disks, a high sensitivity and high throughput inspection system is required to detect defects on the surface of semiconductor wafers or disk substrates. Dark-field optical microscopy with high numerical-aperture detectors in oblique directions is often used to detect weak and anisotropic scattered light from small defects. With conventional high numerical aperture (NA) optics, it is difficult to image the wide field from an oblique direction because of its short depth of focus (DOF). Here, we propose a new optics called “pupil divided optics with magnification correction.” In this optics, a low NA cylindrical lens array is placed on the pupil plane of the high NA objective lens to extend DOF. In this paper, we describe the theory and configuration of the proposed optics. Our simulation and experimental results show the capability for oblique imaging with a wide field.

Microsphere assistance in interference microscopy with high numerical aperture objective lenses

Various attempts have been discussed to overcome the lateral resolution limit and thus to enlarge the fields of application of optical interference microscopy. Microsphere-assisted microscopy and interferometry have proven that the imaging of structures well below Abbe’s resolution limit through near-field assistance is possible if microspheres are placed on the measured surface and utilized as near-field assisting imaging elements. The enhancement of the numerical aperture (NA) by the microspheres as well as photonic nanojets was identified to explain the resolution enhancement, but also whispering gallery modes and evanescent waves are assumed to have an influence. Up to now, to the best of our knowledge, there is no complete understanding of the underlying mechanisms and no model enabling to examine ideal imaging parameters. This contribution is intended to clarify how much the lateral resolution of an already highly resolving Linnik interferometer equipped with 100 × NA 0.9 objective lenses can be further improved by microspheres. Our simulation model developed so far is based on rigorous near-field calculations combined with the diffraction-limited illumination and imaging process in an interference microscope. Here, we extend the model with respect to microsphere-assisted interference microscopy providing a rigorous simulation of the scattered electric field directly above the sphere. Simulation and experimental results will be compared in the three-dimensional spatial frequency domain and discussed in context with ray-tracing computations to achieve an in-depth understanding of the underlying mechanism of resolution enhancement by the microsphere.

Sensitive ultrawideband transparent PVDF-ITO ultrasound detector for optoacoustic microscopy

An ultrasound detection scheme based on a transparent polyvinylidene-fluoride indium-tin-oxide (PVDF-ITO) piezoelectric film is developed for ultrawideband sensitive detection of optoacoustic (OA) signals down to a noise equivalent pressure (NEP) of 8.4 Pa over an effective detection bandwidth extending beyond 30 MHz. The high signal-to-noise ratio and low noise performance are facilitated by employing a two-stage amplifier structure. The PVDF-ITO detector is directly mounted on a commercial high numerical aperture objective lens of a scanning optical resolution OA microscopy system to obtain submicron resolution images without signal averaging, as demonstrated both in phantoms and in vivo measurements in mice. The transparent detection scheme further allows for the OA imaging modality to be easily integrated with other imaging techniques for diverse multi-modal biomedical imaging investigations.

Manipulating the transverse spin angular momentum and Belinfante momentum of spin-polarized light by a tilted stratified medium in optical tweezers

In the recent past, optical tweezers incorporating a stratified medium have been exploited to generate complex translational and rotational dynamics in mesoscopic particles due to the coupling between the spin and orbital angular momentum of the light, generated as a consequence of the tight focusing of light by a high numerical aperture objective lens into the stratified medium. Here, we consider an optical tweezers system with a tilted stratified medium (direction of stratification at an angle with the axis of the incident beam), and show that for input circularly polarized Gaussian beams, the resulting spin-orbit interaction deeply influences the generation of transverse spin angular momentum (TSAM) and Belinfante momentum of light, and allows additional control on their magnitude. Importantly, the TSAM generated in our system consists of both the orthogonal components, which is in sharp contrast to the case of evanescent waves and surface plasmons, where only one of the TSAM components are generated. The broken symmetry due to the tilt ensures that, depending upon the helicity of the input beam, the magnitude of the mutually orthogonal components of the TSAM depend entirely on the tilt angle. This may prove to be an effective handle in exotic spin-controlled manipulation of particles in experiments.

Interference of the scattered vector light fields from two optically levitated nanoparticles.

We experimentally study the interference of dipole scattered light from two optically levitated nanoparticles in vacuum, which present an environment free of particle-substrate interactions. We illuminate the two trapped nanoparticles with a linearly polarized probe beam orthogonal to the propagation of the trapping laser beams. The scattered light from the nanoparticles are collected by a high numerical aperture (NA) objective lens and imaged. The interference fringes from the scattered vector light for the different dipole orientations in image and Fourier space are observed. Especially, the interference fringes of two scattered light fields with polarization vortex show the π shift of the interference fringes between inside and outside the center region of the two nanoparticles in the image space. As far as we know, this is the first experimental observation of the interference of scattered vector light fields from two dipoles in free space. This work also provides a simple and direct method to determine the spatial scales between optically levitated nanoparticles by the interference fringes.

The tight-focusing properties of radially polarized symmetrical power-exponent-phase vortex beam

In this paper, the radially polarized (RP) new kind of power-exponent-phase vortex (NPEPV) beam, with rotationally symmetrical phase structure, was introduced and the tightly focused properties of the RP NPEPV beam passing through a high numerical aperture objective lens were studied numerically. The results show that with the increase of topological charge l, there are multiple intensity points in the focal region, and the number is consistent with the topological charge. In addition, as the power order n increases, the light intensity gradually concentrates on the central optical axis and the surrounding intensity points gradually disappear, which finally presents a Gaussian intensity distribution with the dark cores gradually move away from the optical axis and disappear. These unique properties will have potential applications in particle trapping and laser fabrication, especially for simultaneous trapping of multiple particles and fabrication of chiral microstructures.

Site-resolved imaging of a bosonic Mott insulator of Li7 atoms

We demonstrate a single-site and single-atom-resolved fluorescence imaging of a bosonic Mott insulator of $^7$Li atoms in an optical lattice. The fluorescence images are obtained by implementing Raman sideband cooling on a deep two-dimensional square lattice, where we collect scattered photons with a high numerical aperture objective lens. The square lattice is created by a folded retro-reflected beam configuration that can reach 2.5~mK lattice depth from a single laser source. The lattice beam is elliptically focused to have a large area with deep potential. On average 4,000 photons are collected per atom during 1~s of the Raman sideband cooling, and the imaging fidelity is over 95$\%$ in the central 80$\times$80 lattice sites. As a first step to study correlated quantum phases, we present the site-resolved imaging of a Mott insulator. Tuning the magnetic field near the Feshbach resonance, the scattering length can be increased to 680$a_B$, and we are able to produce a large-sized unity filling Mott insulator with 2,000 atoms at low temperature. Our work provides a stepping stone to further in-depth investigations of intriguing quantum many-body phases in optical lattices.

Generalised adaptive optics method for high-NA aberration-free refocusing in refractive-index-mismatched media.

Phase aberrations are introduced when focusing by a high-numerical aperture (NA) objective lens into refractive-index-mismatched (RIM) media. The axial focus position in these media can be adjusted through either optical remote-focusing or mechanical stage translation. Despite the wide interest in remote-focusing, no generalised control algorithm using Zernike polynomials has been presented that performs independent remote-focusing and RIM correction in combination with mechanical stage translation. In this work, we thoroughly review derivations that model high-NA defocus and RIM aberration. We show through both numerical simulation and experimental results that optical remote-focusing using an adaptive device and mechanical stage translation are not optically equivalent processes, such that one cannot fully compensate for the other without additional aberration compensation. We further establish new orthogonal modes formulated using conventional Zernike modes and discuss its device programming to control high-NA remote-focusing and RIM correction as independent primary modes in combination with mechanical stage translation for aberration-free refocusing. Numerical simulations are performed, and control algorithms are validated experimentally by fabricating graphitic features in diamond using direct laser writing.