In order to solve the problem of the short working distance of the microscope objective in a large aperture, this paper designs a 50× microscope objective with a numerical aperture of 0.75 and a long working distance of 6.2 mm. The objective achieves flat-field and apochromatic for the wavelength range from 436 to 656 nm and maintains high focusing quality in both femtosecond laser (770-790 nm) and YAG laser (1059-1069 nm) ranges. The performance evaluations of the objective show that the resolution reaches 2200 lp/mm, and the modulation transfer function (MTF) indicates that the image quality is close to the diffraction limit within the working ranges. The tolerance analysis reveals that the design meets the requirements for mass production. This paper provides a solution for semiconductor defect detection by combining high resolution with a large working distance.

This work proposes an optimization algorithm in optical design based on the concepts of ergodic and stochastic processes in statistical mechanics. In mixed-variable optimization problems, pseudo-random number and discrete-to-continuous variable conversion dramatically increase the speed at which the system solves for the optimal solution. Pseudo-random numbers are mainly applied in two important steps in the optimization algorithm: determining the combination of glasses involved and the order in which the successive glass parameters are replaced by real glasses. After two series of stochastic processes, the merit function value decreases rapidly along the steepest descent path, and thus the optical system approaches the optimal solution within a very short duration of time. By using the method proposed in this paper, a plan apochromatic objective with a long working distance was optimized, and finally, a high-quality optical system was obtained.

The investigation of deep-sea microorganisms holds immense significance and value in advancing the fields of life sciences, biotechnology, and environmental conservation. However, the current lack of specialized underwater objectives specifically designed for in situ studies of deep-sea microorganisms hampers progress in this area. To address this limitation, we present the design of a multi-band Raman tweezer objective tailored for deep-sea environments. The objective is integrated into a high-pressure chamber capable of withstanding depths up to 1.5 km, enabling in situ microscopic imaging, optical tweezer capture, and Raman detection of deep-sea microorganisms. Through meticulous structural optimization, meticulous material selection, and thorough mechanical analysis of the underwater optical window, the objective exhibits remarkable attributes such as multi-band functionality, extended working distance, and high numerical aperture. Our design yields image quality near the diffraction limit, successfully achieving flat-field and apochromatic performance in each respective wavelength bands. Moreover, the tolerance analysis demonstrates that the full-field root mean square (RMS) wave aberration approaches λ/14, effectively meeting the demands of manufacturing and practical applications. This objective lens constitutes a vital tool for the in situ exploration of deep-sea microorganisms.

Planar super-oscillatory lens (SOL), a far-field subwavelength-focusing diffractive device, holds great potential for achieving sub-diffraction-limit imaging at multiple wavelengths. However, conventional SOL devices suffer from a numerical-aperture-related intrinsic tradeoff among the depth of focus (DoF), chromatic dispersion and focusing spot size. Here, we apply a multi-objective genetic algorithm (GA) optimization approach to design an apochromatic binary-phase SOL having a prolonged DoF, customized working distance (WD), minimized main-lobe size, and suppressed side-lobe intensity. Experimental implementation demonstrates simultaneous focusing of blue, green and red light beams into an optical needle of ~0.5λ in diameter and DOF > 10λ at WD = 428 μm. By integrating this SOL device with a commercial fluorescence microscope, we perform, for the first time, three-dimensional super-resolution multicolor fluorescence imaging of the "unseen" fine structures of neurons. The present study provides not only a practical route to far-field multicolor super-resolution imaging but also a viable approach for constructing imaging systems avoiding complex sample positioning and unfavorable photobleaching.

We present the development of a dual-mode imaging platform that combines optical microscopy with magnetic resonance microscopy. Our microscope is designed to operate inside a 9.4T small animal scanner with the option to use a 72mm bore animal RF coil or different integrated linear micro coils. With a design that minimizes the magnetic distortions near the sample, we achieved a field inhomogeneity of 19 ppb RMS. We further integrated a waveguide in the optical layout for the electromagnetic shielding of the camera, which minimizes the noise increase in the MR and optical images below practical relevance. The optical layout uses an adaptive lens for focusing, 2 × 2 modular combinations of objectives with 0.6mm to 2.3mm field of view and 4 configurable RGBW illumination channels and achieves a plano-apochromatic optical aberration correction with 0.6μm to 2.3μm resolution. We present the design, implementation and characterization of the prototype including the general optical and MR-compatible design strategies, a knife-edge optical characterization and different concurrent imaging demonstrations.

We develop a multimodal imaging platform, combining depth-resolved scattering contrast from spectral-domain optical coherence tomography (SD-OCT) with complementary, non-contact absorption contrast using photoacoustic remote sensing (PARS) microscopy. The system provides a widefield OCT mode using a telecentric scan lens, and a high-resolution, dual-contrast mode using a 0.26 numerical aperture apochromatic objective. An interlaced acquisition approach is used to achieve simultaneous, co-registered imaging. The SD-OCT modality provides a 9.7 µm axial resolution. Comprehensive in vivo imaging of a nude mouse ear is demonstrated, with the SD-OCT scattering intensity revealing dermal morphology, and PARS microscopy providing a map of microvasculature.

In this work, the DC two-photon absorption signal offset (${{\rm DC}_{\rm TPA}}$DCTPA) and the amplitude of the autocorrelation (${{\rm A}_{\rm AC}}$AAC) are measured simultaneously around the focal point of an apochromatic microscope objective using the z-scan autocorrelation technique. The ${{\rm A}_{\rm AC}}$AAC is obtained from the nonlinear sensor response given by the two-photon-absorption, generated in a GaAsP photodiode, for femtosecond laser pulses. We verify that the change in the ${{\rm DC}_{\rm TPA}}$DCTPA signal along $z$z is coincident with the amplitude of the intensity autocorrelation, and that the highest amplitude of the AC is reached at the same position as the highest amplitude of the ${{\rm DC}_{\rm TPA}}$DCTPA signal. The ${{\rm DC}_{\rm TPA}}$DCTPA signal is typically used as a reference for the alignment in a collinear intensity autocorrelator, and we show that it can also be used as a practical procedure to estimate the depth of focus. The ${{\rm DC}_{\rm TPA}}$DCTPA signal measurement allows us to locate the optimum spatial-temporal coupling given by the highest amplitude of the intensity autocorrelation. Additionally, we find a variation in the pulse duration within the same region due to the radially varying group delay dispersion.

Precise machine vision detection for a large object at a finite working distance (WD) requires that the lens has a high resolution for a large field of view (FOV). In this case, the effect of a secondary spectrum on image quality is not negligible. According to the detection requirements, a high resolution apochromatic objective is designed and analyzed. The initial optical structure (IOS) is combined with three segments. Next, the secondary spectrum of the IOS is corrected by replacing glasses using the dispersion vector analysis method based on the Buchdahl dispersion equation. Other aberrations are optimized by the commercial optical design software ZEMAX by properly choosing the optimization function operands. The optimized optical structure (OOS) has an f-number (F/#) of 3.08, a FOV of φ60 mm, a WD of 240 mm, and a modulated transfer function (MTF) of all fields of more than 0.1 at 320 cycles/mm. The design requirements for a nonfluorite material apochromatic objective lens with a large field and high definition for machine vision detection have been achieved.

We demonstrate a method to measure chromatic aberrations of microscope objectives with metallic nanoparticles using white light. Extinction spectra are recorded while scanning a single nanoparticle through a lens's focal plane. We show a direct correlation between the focal wavelength and the longitudinal chromatic focal shift through our analysis of the variations between the scanned extinction spectra at each scan position and the peak extinction over the entire scan. The method has been tested on achromat and apochromat objectives using aluminum disks varying in size from 260-520 nm. Our method is straightforward, robust, low cost, and broadband with a sensitivity suitable for assessing longitudinal chromatic aberrations in high-numerical-aperture apochromatic corrected lenses.

This chapter provides information on how microscopes work and discusses some of the microscope issues to be considered in using a video camera on the microscope. There are two types of microscopes in use today for research in cell biology-the older finite tube-length (typically 160mm mechanical tube length) microscopes and the infinity optics microscopes that are now produced. The objective lens forms a magnified, real image of the specimen at a specific distance from the objective known as the intermediate image plane. All objectives are designed to be used with the specimen at a defined distance from the front lens element of the objective (the working distance) so that the image formed is located at a specific location in the microscope. Infinity optics microscopes differ from the finite tube-length microscopes in that the objectives are designed to project the image of the specimen to infinity and do not, on their own, form a real image of the specimen. Three types of objectives are in common use today-plan achromats, plan apochromats, and plan fluorite lenses. The concept of mounting video cameras on the microscope is also presented in the chapter.

A method of glass selection for the design of optical systems with reduced chromatic aberration is presented. This method is based on the unification of two previously published methods adding new contributions and using a multi-objective approach. This new method makes it possible to select sets of compatible glasses suitable for the design of super-apochromatic optical systems. As an example, we present the selection of compatible glasses and the effective designs for all-refractive optical systems corrected in five spectral bands, with central wavelengths going from 485 nm to 1600 nm.

The possibility and the efficiency of using a single diffractive lens to achromatize and apochromatize micro-objectives with plastic lenses are shown. In addition, recommendations are given on assembling the starting configurations of the objectives and calculating the design parameters required for subsequent optimization. It is also shown that achievable optical performance of achromatic and apochromatic micro-objectives with plastic lenses satisfy the qualifying standards for cell-phone objectives and closed-circuit television (CCTV) cameras.

Double-label techniques were used to concomitantly label olivocochlear neurons and serotoninergic fibers in the bush baby (Otolemur garnettii) brainstem. Light-microscopic examination (using a 100 x plan apochromatic oil-immersion objective) of the sections revealed that serotonin-positive varicosities (presumptive terminal endings) contacted somata and dendrites of neurons belonging to both the lateral and medial olivocochlear neurons near the superior olivary nuclei. These results provide direct evidence that the olivocochlear system (a specific auditory brainstem pathway) receives input from the serotoninergic system (a diffuse reticular brainstem network).

A new method of attaining apochromatic color correction in refractive optical systems has been developed. Common, low-cost optical glasses and abnormal dispersion optical liquids are combined to produce optical systems that have a degree of color correction which previously was possible only by using abnormal dispersion glasses or crystals. In addition to providing dramatic color correction these liquids have good availability, reasonable cost, and low light scatter. Two designs are presented: a glass-liquid apochromatic telescope objective which uses one abnormal dispersion liquid, and an athermal design which uses two liquids. Also presented are the theoretical and experimentally measured optical performance characteristics of these two lenses.