Microscope Objective Research Articles

A three-mode optical thermometer utilizing Cs2NaInCl6: Sb3+/Tb3+ lead-free double perovskite microcrystals for lubricating oil temperature monitoring

Temperature monitoring is essential to maintain proper diesel engine lubricating oil temperature and ensure safe navigation for ships. In contrast to conventional contact thermometers, optical thermometers that operate without physical contact have garnered researchers' attention because of their rapid response, exceptional sensitivity, and capability to measure microscopic objects. Combining multiple thermometric techniques enables self-referenced temperature sensing, thereby improving the accuracy of temperature monitoring signals. Herein, Cs2NaInCl6: Sb3+/Tb3+ microcrystals (MCs) were synthesized to investigate the optical properties and multi-mode temperature detection performances. Upon excitation of 320nm, energy transfer (self-trapped excitons (STEs)→Tb3+) occurs, resulting in blue emission from the STEs and the characteristic emissions from Tb3+. Owing to the superior luminescent properties of Cs2NaInCl6:Sb3+/Tb3+ that vary with temperature, a three-mode optical thermometer has been developed using the ratio of fluorescence intensities (FIR) of STEs and Tb3+ emissions, CIE color coordinates, and the fluorescence lifetime of Tb3+. The reliability of the thermometer has been proven through the practical application of lubricating oil temperature detection, and the measured temperatures closely align with those obtained via infrared thermometry. This work represents a significant step towards developing multi-mode thermometers based on lead-free perovskites.

Pregnancy detection based on blood serum sample Raman spectroscopy.

In this research, women were diagnosed as pregnant based on blood serum samples Raman spectroscopy. Raman spectroscopy is a vibrational technique that provides information on the chemical composition of samples. The Raman techniques have significantly impacted the study of various degenerative diseases, particularly cancer detection, using less invasive methods such as the analysis of blood serum samples. Additionally, these techniques have been used to study the health status of patients, which is often difficult to monitor using conventional techniques. This study obtained serum samples from 12 women diagnosed as pregnant and 11 non-pregnant volunteers (controls). Spectra were collected using a LabRAM HR800 Raman Spectrometer (Horiba Jobin-Yvon) with an 830 nm wavelength laser. For each serum sample, 10 Raman spectra were obtained by focusing the laser, using a microscope objective, on different points of the sample with an exposure time of 40 seconds and an irradiation power of 17 mW. The raw spectra were processed using baseline correction and smoothing to remove noise, fluorescence, and shot noise. Subsequently, the spectra were normalized and analyzed using the multivariate statistical method of Principal Component Analysis (PCA). In these spectra, the characteristic bands of main blood serum biomolecules such as phenylalanine (Phe), tyrosine (Tyr), glutathione, amide III, phospholipid, carotene, and tryptophan (Trp) can be observed. Nevertheless, when analyzing the average spectra of pregnant and non-pregnant women, the main spectral differences were associated with variations in molecules like glutathione, tryptophan (Trp), tyrosine, and phenylalanine, which occur during the first trimesters of pregnancy. This aligns with findings previously reported by other studies. Furthermore, the serum samples from pregnant and non-pregnant patients can be effectively discriminated using PCA applied to the Raman spectra, revealing two distinct clusters in the PCA plot corresponding to each group's status. The results demonstrate that pregnancy can be determined based on blood serum samples Raman spectroscopy with sensitivity and specificity. Although there are very effective devices on the market to determine pregnancy based on the Human Chorionic Gonadotropin (HCG) hormone detection in urine samples, these preliminary results indicate an alternative method known as Raman spectroscopy. On the other hand, the results could also suggest the possibility of carrying out other gynecological or fetal-related studies in women using these Raman techniques.

Micromotor based on single fiber optical vortex tweezer

Optical micromotors are powerful tools for trapping and rotating microparticles in various fields of bio-photonics. Conventionally, optical micromotors are built using bulk optics, such as microscope objectives and SLMs. However, optical fibers provide an attractive alternative, offering a flexible photon platform for optical micromotor applications. In this paper, we present an optical micromotor designed for 3D manipulation and rotation based on a single fiber optical vortex tweezer. A tightly focused vortex beam is excited by preparing a spiral zone plate with an ultrahigh numerical aperture of up to 0.9 at the end facet of a functionalized fiber. The focused vortex beam can optically manipulate and rotate a red blood cell in 3D space far from the fiber end facet. The trapping stiffness in parallel and perpendicular orientations to the fiber axis are measured by stably trapping a standard 3-µm silica bead. The rotational performance is analyzed by rotating a trimer composed of silica beads on a glass slide, demonstrating that the rotational frequency increases with rising optical power and the rotational direction is opposite to the topological charge of the spiral zone plate. The proposed fiber micromotor with its flexible manipulation of microparticle rotation circumvents the need for the precise relative position control of multiple fiber combinations and the use of specialized fibers. The innovations hold promising potential for applications in microfluidic pumping, biopsy, micromanipulation, and other fields.

Trapping light, revealing properties: Laser trapping as a powerful tool for photoluminescence spectroscopy

Laser trapping, a non-contact technique for manipulating microscopic objects, has gained prominence in scientific research. When coupled with fluorescence spectroscopy, it offers a powerful tool for exploring material properties. This study demonstrates the application of laser trapping in the crystallization of MAPbBr3 perovskites and its simultaneous use for photoluminescence imaging. MAPbBr3 perovskites are a class of materials with exceptional optical properties, making them attractive for various optoelectronic applications. However, conventional excitation methods using UV light can lead to phase segregation and mechanical distortion of these materials. Two-photon excitation, on the other hand, offers advantages such as deeper penetration and reduced scattering interference. In this research, we utilize a 1064 nm continuous wave laser for both trapping and excitation purposes. The MAPbBr3 perovskite, with its absorption band ranging from 400 to 540 nm, exhibits two-photon excitation at 532 nm. By focusing the laser beam at the air-solution interface, we successfully crystallize MAPbBr3 perovskites from an unsaturated precursor solution. Simultaneously, the same laser source is used for photoluminescence imaging, allowing for real-time analysis of the crystal's emission properties. This approach eliminates the need for additional excitation sources and simplifies the experimental setup. The combination of laser trapping and two-photon excitation opens up new possibilities for studying perovskite materials. It provides a gentle and non-invasive method for manipulating and characterizing hybrid perovskites materials, paving the way for advancements in various fields such as optoelectronics and energy harvesting.

Imaging of Live Cells by Digital Holographic Microscopy

Imaging of microscopic objects is of fundamental importance, especially in life sciences. Recent fast progress in electronic detection and control, numerical computation, and digital image processing, has been crucial in advancing modern microscopy. Digital holography is a new field in three-dimensional imaging. Digital reconstruction of a hologram offers the remarkable capability to refocus at different depths inside a transparent or semi-transparent object. Thus, this technique is very suitable for biological cell studies in vivo and could have many biomedical and biological applications. A comprehensive review of the research carried out in the area of digital holographic microscopy (DHM) for live-cell imaging is presented. The novel microscopic technique is non-destructive and label-free and offers unmatched imaging capabilities for biological and bio-medical applications. It is also suitable for imaging and modelling of key metabolic processes in living cells, microbial communities or multicellular plant tissues. Live-cell imaging by DHM allows investigation of the dynamic processes underlying the function and morphology of cells. Future applications of DHM can include real-time cell monitoring in response to clinically relevant compounds. The effect of drugs on migration, proliferation, and apoptosis of abnormal cells is an emerging field of this novel microscopic technique.

A perspective on guided electrophoretic transport of particles in liquid crystals

Nonlinear electrophoresis in complex fluids like nematic liquid crystals provides new pathways toward achieving precisely controlled motion and assembly of microscopic objects. The nematic host introduces a paradigm shift in the mechanism of electrophoretic transport by generating unbalanced electro-osmotic flows around the colloidal particle due to symmetry breaking of the medium caused by the induced topological defects. Rationally designed particles, which induce various types of defects and asymmetries, provide new opportunities in this regard. In this Perspective article, we discuss how the asymmetry in the shape and interfacial properties help in piloting the particles using an AC electric field. Finally, we propose some feasible strategies to achieve navigational control using magnetic and photo-responsive particles, guided by orthogonal electric, magnetic fields, and light, respectively.

Development and Testing of a Dual-Driven Piezoelectric Microgripper with High Amplification Ratio for Cell Micromanipulation

Cell micromanipulation is an important technique in the field of biomedical engineering. Microgrippers play a crucial role in connecting macroscopic and microscopic objects in micromanipulation systems. However, since the operated biological cells are deformable, vulnerable, and typically distributed in sizes ranging from micrometers to millimeters, it poses a huge challenge to microgripper performance. To solve this problem, this paper develops a dual-driven piezoelectric microgripper with a high displacement amplification ratio, large stroke, and parallel gripping. By adopting modular configuration, three kinds of flexure-based mechanisms, including the lever mechanism, Scott–Russell mechanism, and parallelogram mechanism are connected in series to realize three-stage amplification, which effectively makes up for the shortage of small output displacement of the piezoelectric actuator. At the same time, the use of the parallelogram mechanism also isolates the parasitic rotation movement, and realizes the parallel movement of the gripping jaws. In addition, the kinematics, statics, and dynamics models of the microgripper are established by using the pseudo-rigid body and Lagrange methods, and the key geometric parameters are also optimized. Finite element simulation and experimental tests verify the effectiveness of the developed microgripper. The results show that the developed microgripper allows an amplification ratio of 46.4, a clamping stroke of 2180 μm, and a natural frequency of 203.1 Hz. Based on the developed microgripper, the nondestructive micromanipulation of zebrafish embryos is successfully realized.

Photomechanical Crystals as Light-Activated Organic Soft Microrobots.

In the field of materials science, dynamic molecular crystals have attracted significant attention as a novel class of energy-transducing materials. However, their development into becoming fully functional actuators remains somewhat limited. This study focuses on one family of dynamic crystalline materials and delves into exploring the efficiency of conversion of light energy to mechanical work. A simple setup is designed to determine a set of performance indices of anthracene-based crystals as an exemplary class of dynamic molecular crystals. The ability of these crystals to reversibly bend due to dimerization is realistically assessed from the perspective of the envisaged soft robotics applications, where wireless photomechanical grippers manipulate and assemble microscopic objects driven and controlled by light instead of lines and motors. The approach described here not only guides the quantification of responsive molecular crystals' actuation potential but also aims to attract an interdisciplinary interest to further develop this class of materials into controllable all-organic actuating elements to be used in microrobotics for engineering or biomedicine.

Lensless Mueller holographic microscopy with robust noise reduction for multiplane polarization imaging

Lensless holographic microscopy has emerged as a powerful and cost-effective tool for computational imaging, offering high resolution over a large field of view, beneficial for various biological applications. However, conventional approaches can struggle with contrast and accurate visualization of diverse components over the samples, which can directly affect the diagnostic precision of the techniques. Mueller imaging, while offering detailed, stain-free observations of polarized light responses in samples, often has a limited field of view and single plane information. This is due to the use of high NA microscope objectives and generally complex hardware setups, thus narrowing its practical effectiveness. This work introduces a Lensless Mueller Holographic Microscopy (LMHM) system that overcomes these limitations, enabling large field of view, volumetric multi-layer imaging, and Mueller matrix computation using in-line lens-free holography setup. The proposed system provides precision visualization of polarization information in samples, offering high-quality features due to the incorporation of a numerical multi-height Gerchberg-Saxton reconstruction algorithm with additional complex field filtering and a physical rotating diffuser. The proposed LMHM framework is validated with a calibrated USAF 1951 birefringent test target. A multiplane sample containing cloth fiber is utilized to study the LMHM capabilities of imaging volumetric samples. Finally, the LMHM is used to analyze two mice’s brain slices, effectively showcasing this organ’s anatomy. Among other structures in the brain, the proposed method easily allows the visualization of, e.g., the corpus callosum. These results constitute a proof-of-concept evaluation for bioimaging applications.

Characterization of a levitated sub-milligram ferromagnetic cube in a planar alternating-current magnetic Paul trap

Microscopic levitated objects are a promising platform for inertial sensing, testing gravity at small scales, optomechanics in the quantum regime, and large-mass superpositions. However, existing levitation techniques harnessing optical and electrical fields suffer from noise induced by elevated internal temperatures and charge noise, respectively. Meissner-based magnetic levitation circumvents both sources of decoherence but requires cryogenic environments. Here, we characterize a sub-milligram ferromagnetic cube levitated in an alternating-current planar magnetic Paul trap at room temperature. We show behavior in line with the Mathieu equations and quality factors of up to 2500 for the librational modes. Besides technological sensing applications, this technique sets out a path for megahertz librational modes in the micrometer-sized particle limit and can be extended by implementing superconducting traps in cryogenic environments, allowing for magnetic coupling to superconducting circuits and spin-based quantum systems.

In silico full-angle high-dynamic range scattering of microscopic objects exploiting holotomography.

Accurate optical characterization of microscopic objects is crucial in academic research, product development, and clinical diagnosis. We present a method for obtaining full and high-dynamic range, angle-resolved light scattering attributes of microparticles, enabling distinction of variations in both overall morphology and detailed internal structures. This method overcomes previous limitations in observable scattering angles and dynamic range of signals through computationally assisted three-dimensional holotomography. This advancement is significant for particles spanning tens of wavelengths, such as human erythrocytes, which have historically posed measurement challenges due to faint side-scattering signals indicative of their complex interiors. Our technique addresses three key challenges in optical side-scattering analysis: limited observational angular range, reliance on simplified computational models, and low signal-to-noise ratios in both experimental and computational evaluations. We incorporate three-dimensional tomographic complex refractive index data from Fourier-transform light scattering into a tailored finite-difference time-domain simulation space. This approach facilitates precise near-to-far-field transformations. The process yields complete full-angle scattering phase functions, crucial for particles like Plasmodium falciparum-parasitized erythrocytes, predominantly involved in forward scattering. The resultant scattering data exhibit an extreme dynamic range exceeding 100 dB at various incident angles of a He-Ne laser. These findings have the potential to develop point-of-care, cost-effective, and rapid malaria diagnostic tools, inspiring further clinical and research applications in microparticle scattering.

Deep learning and defocus imaging for determination of three-dimensional position and orientation of microscopic objects

We present a method to determine the three-dimensional position and orientation of microscopic, non-spherical objects in microfluidic and laboratory-on-a-chip systems observed through conventional optical microscopes. The method is based on the combination of the General Defocusing Particle Tracking technique [Barnkob et al., “General defocusing particle tracking,” Lab Chip 15, 3556–3560 (2015)] and deep learning. It requires minimal input from the user, is suitable for real-time applications, and can be applied to any microscopic object with an approximately ellipsoidal shape, such as unicellular swimming organisms, red blood cells, or spheroidal colloids. The main challenge is linked to the construction of suitable training datasets for the neural network. We provide a procedure generally valid for active microswimmers and discuss possible strategies for other types of objects. An implementation using the Visual Geometry Group convolutional neural network (VGG-16) is presented and tested on synthetic images with different backgrounds and noise levels. The same implementation is used to track the position and orientation of different specimens of the heterotrophic ciliate Euplotes Vannus in free-swimming motion. The measurements were performed with a 10 × objective over a depth of 800 μm with an average estimated uncertainty in the orientation angles of 9.0%.

Switchable optical trapping and manipulation enabled by polarization-modulated multifunctional phase-change metasurfaces

Optical trapping, a cutting-edge methodology, is pivotal for contactlessly controlling and exploring microscopic objects. However, it encounters formidable challenges such as multiparticle trapping, flexible control, and seamless integration. Here, we employ a polarization-modulated multi-foci technique for versatile nanoparticle trapping using multifunctional metasurfaces relying on geometric phase. Numerical simulations demonstrate the generation of two focused spots with orthogonal polarization distributions through our metasurfaces when illuminated with linearly polarized light, with their polarization distributions be interchanged by orthogonally switching the incident polarizations. We extend this design to an array of multi-foci metasurface tweezers modulated by polarization, highlighting the versatility and robustness of our approach. Furthermore, we demonstrate the simultaneous generation of two distinct focusing cylindrical vector beams using a monolayer metasurface, showcasing the two vector beams possess the interchange ability of their polarization distributions. By leveraging the Maxwell stress tensor, we assess the distinct contributions of the focused beams to longitudinal and transverse optical forces on SiO2 spheres, validating diverse trapping and manipulation behaviors for nanoparticles with the proposed metasurface designs. By manipulating the phase states of Sb2S3 nanopillars, binary-switchable optical trapping and manipulation are facilitated for all proposed metasurface tweezers. Our work underscores the efficacy of polarization-modulation multifunctional metasurface tweezers in consolidating multiple trapping tasks into a single device, paving the way for innovative lab-on-a-chip optical trapping applications in biophysics, nanotechnology, and photonics.

Moving microscopic objects with self-disassembly.

Moving microscopic objects with self-disassembly.

Design of a catadioptric mid-wave infrared telecentric microscope with high numerical aperture and long working distance

Infrared microscopy is widely used in the field of thermal analysis. This paper proposes a design method for a catadioptric mid-wave infrared telecentric microscope system that solves the problem of traditional microscopy systems being unable to simultaneously meet high numerical aperture, long working distance and large magnification requirements. The system is composed of a microscope objective and a tubelens. The objective uses an improved Schwarzschild structure and is connected to the tubelens through an externally located pupil. The system works in the wavelength range of 3.7 μm–4.8 μm, achieves a 0.82 numerical aperture, a 15.5 mm working distance, an 8× magnification and a 2.75 μm object-side resolution. The image size of the detector is 17um, and the MTF of the microscope system at the detector cutoff frequency is greater than 0.2, which is close to the diffraction limit. Finally, tolerance analysis and athermal analysis are conducted on the system, and the results indicate that the system possesses good manufacturability and thermal stability.

The bacterial flagellum as an object for optical trapping.

This letter considers the possibility of using the optical trap to study the structure and function of the microbial flagellum. The structure of the flagellum of a typical gram-negative bacterium is described in brief. A standard mathematical model based on the principle of superposition is used to describe the movement of an ellipsoidal microbial cell in a liquid medium. The basic principles of optical trapping based on the combined action of the light pressure and the gradient force are briefly clarified. Several problems related to thermal damage of living microscopic objects when the latter gets to the focus of a laser beam are shortly discussed. It is shown that the probability of cell damage depends nonlinearly on the wavelength of laser radiation. Finally, the model systems that would make it possible to study flagella of the free bacteria and the ones anchored or tethered on the surface of a solid material are discussed in detail.

Off-axis reflective microscope objective with a centimeter scale field of view and micron resolution.

Microscope objectives with wide field-of-view (FOV) and high resolution are urgently needed for the frontier research in life sciences. However, traditional transmission microscope objectives typically have a narrow FOV and severe chromatic aberration. A new off-axis reflective microscope objective with a centimeter scale FOV and micron resolution is proposed in this paper. This objective, with its simple structure, can operate over a wide wavelength range. A design method for a wide FOV optical system is presented, which can eliminate the obstruction of the rays and control the intermediate image plane. Using this method, we design a novel off-axis four-mirror microscope objective with a FOV of 10 mm × 1.5 mm and a numerical aperture of 0.33.

Curved Surfaces Induce Metachronal Motion of Microscopic Magnetic Cilia.

Cilia are hair-like organelles present on cell surfaces. They often exhibit a collective wave-like motion that can enhance fluid or particle transportation function, known as metachronal motion. Inspired by nature, researchers have developed artificial cilia capable of inducing metachronal motion, especially magnetic actuation. However, current methods remain intricate, requiring either control of the magnetic or geometrical properties of individual cilia or the generation of a complex magnetic field. In this paper, we present a novel elegant method that eliminates these complexities and induces metachronal motion of arrays of identical microscopic magnetic artificial cilia by applying a simple rotating uniform magnetic field. The key idea of our method is to place arrays of cilia on surfaces with a specially designed curvature. This results in consecutive cilia experiencing different magnetic field directions at each point in time, inducing a phase lag in their motion, thereby causing collective wave-like motion. Moreover, by tuning the surface curvature profile, we can achieve diverse metachronal patterns analogous to symplectic and antiplectic metachronal motion observed in nature, and we can even devise novel combinations thereof. Furthermore, we characterize the local flow patterns generated by the motion of the cilia, revealing the formation of vortical patterns. Our novel approach simplifies the realization of miniaturized metachronal motion in microfluidic systems and opens the possibility of controlling flow pattern generation and transportation, opening avenues for applications such as lab-on-a-chip technologies, organ-on-a-chip platforms, and microscopic object propulsion.

Large-scale microscope with improved resolution using SRGAN

Due to the limited Space-Bandwidth Product (SBP) of optical microscopy system, the Numerical Aperture (NA) and Field of View (FOV)/ Depth of Field (DOF) of conventional optical microscopes are mutually restricted. In this paper, we propose a 20× large-scale (large FOV&DOF) microscope objective lens which is enhanced by deep neural network to achieve the improved resolution close to a 40× Olympus objective lens. Based on training a Super-resolution Generative Adversarial Network (SRGAN) model to transform low-resolution images into super-resolved ones, the resolution of input images captured by low magnification objective is greatly improved without any loss of FOV. And the network output images have better performance on DOF. We also compared SRGAN with two other deep-learning based methods to prove its superiority. Such large-scale microscope with improved resolution could serve to democratize super-resolution imaging.

Nanoalignment by critical Casimir torques

The manipulation of microscopic objects requires precise and controllable forces and torques. Recent advances have led to the use of critical Casimir forces as a powerful tool, which can be finely tuned through the temperature of the environment and the chemical properties of the involved objects. For example, these forces have been used to self-organize ensembles of particles and to counteract stiction caused by Casimir-Liftshitz forces. However, until now, the potential of critical Casimir torques has been largely unexplored. Here, we demonstrate that critical Casimir torques can efficiently control the alignment of microscopic objects on nanopatterned substrates. We show experimentally and corroborate with theoretical calculations and Monte Carlo simulations that circular patterns on a substrate can stabilize the position and orientation of microscopic disks. By making the patterns elliptical, such microdisks can be subject to a torque which flips them upright while simultaneously allowing for more accurate control of the microdisk position. More complex patterns can selectively trap 2D-chiral particles and generate particle motion similar to non-equilibrium Brownian ratchets. These findings provide new opportunities for nanotechnological applications requiring precise positioning and orientation of microscopic objects.