Highly-Adaptable Optothermal Nanotweezers for Trapping, Sorting, and Assembling across Diverse Nanoparticles.

Optical manipulation of various kinds of nanoparticles is vital in biomedical engineering. However, classical optical approaches demand higher laser power and are constrained by diffraction limits, necessitating tailored trapping schemes for specific nanoparticles. They lack a universal and biocompatible tool to manipulate nanoparticles of diverse sizes, charges, and materials. Through precise modulation of diffusiophoresis and thermo-osmotic flows in the boundary layer of an optothermal-responsive gold film, highly adaptable optothermal nanotweezers (HAONTs) capable of manipulating a single nanoparticle as small as sub-10 nm are designed. Additionally, a novel optothermal doughnut-shaped vortex (DSV) trapping strategy is introduced, enabling a new mode of physical interaction between cells and nanoparticles. Furthermore, this versatile approach allows for the manipulation of nanoparticles in organic, inorganic, and biological forms. It also offers versatile function modes such as trapping, sorting, and assembling of nanoparticles. It is believed that this approach holds the potential to be a valuable tool in fields such as synthetic biology, optofluidics, nanophotonics, and colloidal science.

CRISPR-powered optothermal nanotweezers: Diverse bio-nanoparticle manipulation and single nucleotide identification.

Optothermal nanotweezers have emerged as an innovative optical manipulation technique in the past decade, which revolutionized classical optical manipulation by efficiently capturing a broader range of nanoparticles. However, the optothermal temperature field was merely employed for in-situ manipulation of nanoparticles, its potential for identifying bio-nanoparticles remains largely untapped. Hence, based on the synergistic effect of optothermal manipulation and CRIPSR-based bio-detection, we developed CRISPR-powered optothermal nanotweezers (CRONT). Specifically, by harnessing diffusiophoresis and thermo-osmotic flows near the substrate upon optothermal excitation, we successfully trapped and enriched DNA functionalized gold nanoparticles, CRISPR-associated proteins, as well as DNA strands. Remarkably, we built an optothermal scheme for enhancing CRISPR-based single-nucleotide polymorphism (SNP) detection at single molecule level, while also introducing a novel CRISPR methodology for observing nucleotide cleavage. Therefore, this innovative approach has endowed optical tweezers with DNA identification ability in aqueous solution which was unattainable before. With its high specificity and feasibility for in-situ bio-nanoparticle manipulation and identification, CRONT will become a universal tool in point-of-care diagnosis, biophotonics, and bio-nanotechnology.

Multi-color two-photon scanning structured illumination microscopy imaging of live cells.

Multi-color two-photon microscopy imaging of live cells is essential in biology. However, the limited diffraction resolution of conventional two-photon microscopy restricts its application to subcellular organelle imaging. Recently, we developed a laser scanning two-photon non-linear structured illumination microscope (2P-NLSIM), whose resolution improved three-fold. However, its ability to image polychromatic live cells under low excitation power has not been verified. Here, to improve the reconstruction super-resolution image quality under low excitation power, we increased the image modulation depth by multiplying the raw images with the reference fringe patterns in the reconstruction process. Simultaneously, we optimized the 2P-NLSIM system to image live cells, including the excitation power, imaging speed, and field of view. The proposed system could provide a new imaging tool for live cells.

Corrigendum: High-Sensitive Surface Plasmon Resonance Imaging Biosensor Based on Dual-Wavelength Differential Method.

[This corrects the article DOI: 10.3389/fchem.2021.801355.].

Optothermophoretic flipping method for biomolecule interaction enhancement.

The widely used surface-based biomolecule sensing scheme has greatly facilitated the investigation of protein-protein interactions in lab-on-a-chip microfluidic systems. However, in most biosensing schemes, the interactions are driven in a passive way: The overall sensing time and sensitivity are totally dependent on the Brownian diffusion process, which has greatly hindered their efficiency, especially at low concentration levels or single-molecule analysis. To break this limitation, we developed an all-optical active method termed optothermophoretic flipping (OTF). It is the first temporal modulated method that biomolecules were enriched and pushed to their counterparts for effective contact via a flipped thermophoresis. As a proof-of-concept experiment, we tested its performance via antibody-antigen binding on a surface plasmon resonance imaging (SPRi) platform. Compared with the interaction solely based on Brownian diffusion, we achieved a 23.6-fold sensitivity increment in biomolecule interactions sensing. This method has opened new opportunities for various biosensing platforms that require high-sensitivity in colloidal sciences and biochemical studies.

High-Sensitive Surface Plasmon Resonance Imaging Biosensor Based on Dual-Wavelength Differential Method.

Intensity interrogation surface plasmon resonance (ISPR) sensing has a simple schematic design and is the most widely used surface plasmon resonance technology at present. However, it has relatively low sensitivity, especially for ISPR imaging (ISPRi). In this paper, a new technique for the real-time monitoring of biomolecule binding on sensor surfaces via ISPRi detection is described. The technique is based on the interrogation of the differential value of two intensities at two specific wavelengths from the reflected light spectrum. In addition, we also optimized the selection of dual-wavelength parameters under different circumstances to achieve the highest sensitivity. The new technique achieved a refractive index resolution (RIR) of 2.24 × 10-6 RIU, which is far beyond that of traditional ISPRi technique. Moreover, our new ISPRi technique also realized the real-time detection of high-throughput biomolecular binding. This study is expected to promote the development of faster and more accurate SPRi technologies.

Phase interrogation surface plasmon resonance hyperspectral imaging sensor for multi-channel high-throughput detection.

Phase interrogation surface plasmon resonance (SPR) imaging is, in principle, suitable in multiple samples and high-throughput detection, but the refractive index difference of various samples can be largely varied, while the dynamic range of phase interrogation SPR is narrow. So it is difficult to perform multi-sample detection in phase interrogation mode. In this paper, we successfully designed a multi-channel phase interrogation detection SPR imaging sensing scheme based on a common optical interference path between p- and s-polarized light without using any mechanical moving components. The fixed optical path difference between p- and s-polarized light is introduced by a birefringence crystal to produce sinusoidal spectral interference fringes. We adopted a time-division-multiplexing peak-finding algorithm to track the resonance wavelength so that the detection range can cover every channel. The phase values which carry the high sensitivity signal of the corresponding samples are calculated by the iterative parameter scanning cross-correlation algorithm.

Ultrafast Surface Plasmon Resonance Imaging Sensor via the High-Precision Four-Parameter-Based Spectral Curve Readjusting Method.

A variety of surface plasmon resonance (SPR) sensing devices have been extensively used in biochemical detection for their characteristics of label-free, highly sensitive, and faster detecting. Among them, the spectrum-based SPR sensing devices have offered us great advantages in high-throughput sensing due to their large dynamic range and the possibility of detection resolution similar to that offered by angle interrogation. This paper demonstrates a spectrum-based SPR imaging sensing system with fast wavelength scanning capability achieved by an acousto-optic tunable filter (AOTF) and a low-cost and speckle-free halogen lamp implemented as the SPR excitation source. Especially, we developed a novel four-parameter-based spectral curve readjusting (4-PSCR) method for data processing, which offered us a faster and more accurate spectral data curve fitting process than the traditional polynomial fitting method. With the configuration, we have also conducted an SPR high-throughput detection of the novel coronavirus (COVID-19) spike protein, proving its application possibility in the screening of COVID-19 with high accuracy. We believe that the higher sensitivity and accuracy of the system have made it readily used in biochemical imaging and detecting applications.

High-throughput imaging surface plasmon resonance biosensing based on ultrafast two-point spectral-dip tracking scheme.

Wavelength interrogation surface plasmon resonance imaging (λSPRi) has potential in detecting 2-dimensional (2D) sensor array sites, but the resonance wavelength imaging rate limits the application of detecting biomolecular binding process in real time. In this paper, we have successfully demonstrated an ultrafast λSPRi biosensor system. The key feature is a two-point tracking algorithm that drives the liquid crystal tunable filter (LCTF) to achieve fast-tracking of the resonance wavelength movement caused by the binding of target molecules with the probe molecules on the sensing surface. The resonance wavelength measurement time is within 0.25s. To date, this is the fastest speed ever reported in λSPRi. Experiment results show that the sensitivity and dynamic are 2.4 × 10-6 RIU and 4.6 × 10-2 RIU, respectively. In addition, we have also demonstrated that the system has the capability of performing fast high-throughput detection of biomolecular interactions, which confirms that this fast real-time detecting approach is most suitable for high-throughput and label-free biosensing applications.

In Vivo-Like Morphology of Intercalated Discs Achieved in a Neonatal Cardiomyocyte Culture Model.

In vitro cultures to be used in various analytical investigations of cardiomyocyte (CM) growth and function for enhancing insight into physiological and pathological mechanisms should closely express in vivo morphology. The aim of the studies is to explore how to use microfabrication and physical-cue-addition techniques to establish a neonatal rat CM culture model that expresses an end-to-end connected rod shape with in vivo-like intercalated discs (ICDs). Freshly isolated neonatal rat CMs were cultured on microgrooved polydimethylsiloxane substrate. Cell alignment and ICD orientation were evaluated using confocal fluorescence and transmission electron microscopy under various combinations of different culture conditions. Cyclic stretch and blebbistatin tests were conducted to explore mechanical and electrical effects. Laboratory-made MATLAB software was developed to quantify cell alignment and ICD orientation. Our results demonstrate that the mechanical effect associated with the electrical stimulation may contribute to step-like ICD formation viewed from the top. In addition, our study reveals that a suspended elastic substrate that was slack with scattered folds, not taut, enabled CM contraction of equal strength on both apical and basal cell surfaces, allowing the cultured CMs to express a three-dimensional rod shape with disc-like ICDs viewed cross-sectionally. Impact statement In this article, we describe how the tugging forces generated by cardiomyocytes (CMs) facilitate the formation of the morphology of the intercalated discs (ICDs) to achieve mechanoelectrical coupling between CMs. Correspondingly, we report experimental techniques we developed to enable the in vivo-like behavior of the tugging forces to support the development of in vivo-like morphology in ICDs. These techniques will enhance insight into physiological and pathological mechanisms related to the development of tissue-engineered cardiac constructs in various analytical investigations of CM growth and function.

Phase interrogation SPR sensing based on white light polarized interference for wide dynamic detection range.

A phase surface plasmon resonance (SPR) sensing technology based on white light polarized interference in common-path geometry is reported. A halogen lamp is used as the excitation source of the SPR sensor. The fixed optical path difference (OPD) between p- and s-polarized light is introduced by a birefringence crystal to produce sinusoidal spectral interference fringes. The SPR phase is accurately extracted from the interference fringes using a novel iterative parameter-scanning cross-correlation algorithm. The dynamic detection range is expanded by tracking the best SPR wavelength, which is identified using a window Fourier algorithm. The experimental results show that the sensitivity of this SPR system was 1.3 × 10-7 RIU, and the dynamic detection range was 0.029 RIU. This sensor, not only simple to implement and cost efficient, requires no modulators.

Light field 3D measurement using unfocused plenoptic cameras.

This Letter reports a novel method to establish the metric relationship of depth value between object space and image space for unfocused plenoptic cameras. A three-dimensional (3D) measurement system was introduced to precisely construct benchmarks and matching features to compute the metric depths in the object space and the corresponding depth values in the image space for metric calibration. After metric calibration, precise measurement of the depth dimension was possible. Furthermore, with the aid of metric spatio-angular parameters determined via light field ray calibration, transverse dimensions were computed from the measured depth, realizing light field 3D measurement for unfocused plenoptic cameras. Finally, we experimentally performed accuracy analysis of the proposed method with depth measurement precision of 0.5 mm in a depth range of 300 mm, which illuminated potential applications of unfocused plenoptic cameras in the field of 3D measurement.

Wavelength-Scanning SPR Imaging Sensors Based on an Acousto-Optic Tunable Filter and a White Light Laser.

A fast surface plasmon resonance (SPR) imaging biosensor system based on wavelength interrogation using an acousto-optic tunable filter (AOTF) and a white light laser is presented. The system combines the merits of a wide-dynamic detection range and high sensitivity offered by the spectral approach with multiplexed high-throughput data collection and a two-dimensional (2D) biosensor array. The key feature is the use of AOTF to realize wavelength scan from a white laser source and thus to achieve fast tracking of the SPR dip movement caused by target molecules binding to the sensor surface. Experimental results show that the system is capable of completing a SPR dip measurement within 0.35 s. To the best of our knowledge, this is the fastest time ever reported in the literature for imaging spectral interrogation. Based on a spectral window with a width of approximately 100 nm, a dynamic detection range and resolution of 4.63 × 10-2 refractive index unit (RIU) and 1.27 × 10-6 RIU achieved in a 2D-array sensor is reported here. The spectral SPR imaging sensor scheme has the capability of performing fast high-throughput detection of biomolecular interactions from 2D sensor arrays. The design has no mechanical moving parts, thus making the scheme completely solid-state.

High-throughput imaging surface plasmon resonance biosensing based on an adaptive spectral-dip tracking scheme.

Imaging-based spectral surface plasmon resonance (λSPR) biosensing is predominantly limited by data throughput because of the multiplied data capacity emerging from 2-dimensional sensor array sites and the many data points required to produce an accurate measurement of the absorption dip. Here we present an adaptive feedback approach to address the data throughput issue in λSPR biosensing. A feedback loop constantly tracks the dip location while target-molecule binding occurs at the sensor surface. An adaptive window is then imposed to reduce the number of data points that each pixel has to capture without compromising measurement accuracy. Rapid wavelength scanning is performed with a liquid crystal tunable filter (LCTF). With the use of a feedback loop, our demonstration system can produce a dip measurement within 700ms, thus confirming that the reported λSPR approach is most suitable for real-time micro-array label-free biosensing applications.

Fast spectral surface plasmon resonance imaging sensor for real-time high-throughput detection of biomolecular interactions.

A fast surface plasmon resonance (SPR) imaging biosensor system based on wavelength interrogation using a liquid crystal tunable filter (LCTF) is presented. The system combines the merits of wide-dynamic detection range offered by the spectral approach and multiplexed high-throughput data collection with a two-dimensional (2-D) biosensor array. The key feature of the reported scheme is a feedback loop that drives the LCTF to achieve fast tracking of the SPR dip movement caused by the binding of target molecules to the sensor surface. Experimental results show that the system is capable of completing an SPR dip measurement within 4 s. Based on using a spectral window of about 100 nm, the experimental dynamic detection range and refractive index resolution are 4.63×10?2??RIU and 5.87×10?6??RIU, respectively. As also demonstrated herein using 2-D microsensor arrays, among the spectral SPR sensors, the reported system is most suitable for multiplexed label-free biosensing applications.

Improved Rotating Kernel Transformation Based Contourlet Domain Image Denoising Framework.

A contourlet domain image denoising framework based on a novel Improved Rotating Kernel Transformation is proposed, where the difference of subbands in contourlet domain is taken into account. In detail: (1). A novel Improved Rotating Kernel Transformation (IRKT) is proposed to calculate the direction statistic of the image; The validity of the IRKT is verified by the corresponding extracted edge information comparing with the state-of-the-art edge detection algorithm. (2). The direction statistic represents the difference between subbands and is introduced to the threshold function based contourlet domain denoising approaches in the form of weights to get the novel framework. The proposed framework is utilized to improve the contourlet soft-thresholding (CTSoft) and contourlet bivariate-thresholding (CTB) algorithms. The denoising results on the conventional testing images and the Optical Coherence Tomography (OCT) medical images show that the proposed methods improve the existing contourlet based thresholding denoising algorithm, especially for the medical images.

Microfabrication, surface modification, and laser guidance techniques to create a neuron biochip.

In this report we illustrate our application of soft lithography-based microfabrication, surface modification, and our unique laser cell-patterning system toward the creation of neuron biochips. We deposited individual forebrain neurons from Day 7 embryonic chicks into two rows of eight in a silicon microstructure aligned over a microelectrode array (MEA). The polydimethylsiloxane (PDMS) membrane with microstructures to confine cells and guide network connectivity was aligned to the electrodes of a MEA. Both the MEA and the PDMS membrane were treated with O2 plasma, Poly-L-Lysine, and Laminin to aid in cell attachment and survival. The primary advantage of our process is that it is quicker and simpler than previous cell-placement methods and may make highly defined neuronal network biochips more practical.

Dimensionless parameters for the design of optical traps and laser guidance systems.

Optical traps are routinely used for the manipulation of neutral particles. However, optical trap design is limited by the lack of an accurate theory. The generalized Lorenz-Mie theory (GLMT) solves the scattering problem for arbitrary particle size and predicts radial forces accurately. Here we show that the GLMT predicts the observed radial and axial forces in a variety of optical manipulators. We also present a dimensionless parameter beta for the prediction of axial forces. Coupled with our correlation for radial escape forces, we now have a set of two simple correlations for the practical design of radiation-force-based systems.