Cuboids Prevail When Unraveling the Influence of Microchip Geometry on Macrophage Interactions and Metabolic Responses.

Drug delivery advances rely on using nano- and microsized carriers to transfer therapeutic molecules, although challenges persist in increasing the availability of new and even approved pharmaceutical products. Particle shape, a critical determinant in how these carriers distribute within the body after administration, raises opportunities of using, for instance, micrometer-sized nonspherical particles for vascular targeting and thereby creating new prospects for precise drug delivery to specific targeted areas. The versatility of polycrystalline silicon microfabrication allows for significant variation in the size and shape of microchips, and so, in the current work, photolithography was employed to create differently shaped polysilicon microchips, including cuboids, cubes, bars, and cylinders, to explore the influence of particle shape on cellular interactions. These microchips with different shapes and lateral dimensions, accounting for surface areas in the range of ca. 15 to 120 µm2 and corresponding total volumes of 0.4 to 27 µm3, serve as ideal models for investigating their interactions with macrophages with diameters of ca. 20 µm. Side-scattering imaging flow cytometry was employed for studying the interaction of label-free prepared microchips with RAW 264.7 macrophages. Using a dose of 3 microchips per cell, results show that cuboids exhibit the highest cellular association (ca. 25%) and uptake (ca. 20%), suggesting their potential as efficient carriers for targeted drug delivery to macrophages. Conversely, similarly sized cylinders and bar-shaped microchips exhibit lower uptakes of about 8% and about 6%, respectively, indicating potential benefits in evading macrophage recognition. On average, 1-1.5 microchips were internalized, and ca. 1 microchip was surface-bound per cell, with cuboids showing the higher values overall. Macrophages respond to microchips by increasing their metabolic activity and releasing low levels of intracellular enzymes, indicating reduced toxicity. Interestingly, increasing the particle dose enhances macrophage metabolic activity without significantly affecting enzyme release.

Surface-Enhanced Raman Scattering Imaging Assisted by Machine Learning Analysis: Unveiling Pesticide Molecule Permeation in Crop Tissues.

Surface-enhanced Raman scattering (SERS) imaging technology faces significant technical bottlenecks in ensuring balanced spatial resolution, preventing image bias induced by substrate heterogeneity, accurate quantitative analysis, and substrate preparation that enhances Raman signal strength on a global scale. To systematically solve these problems, artificial intelligence techniques are applied to analyze the signals of pesticides based on 3D and dynamic SERS imaging. Utilizing perovskite/silver nanoparticles composites (CaTiO3/Ag@BONPs) as enhanced substrates, enabling it not only to cleanse pesticide residues from the surface to pulp of fruits and vegetables, but also to investigate the penetration dynamics of an array of pesticides (chlorpyrifos, thiabendazole, thiram, and acetamiprid). The findings challenge existing paradigms, unveiling a previously unnoticed weakening process during pesticide invasion and revealing the surprising permeability of non-systemic pesticides. Of particular note is easy to overlook that the combined application of pesticides can inadvertently intensify their invasive capacity due to pesticide interactions. The innovative study delves into the realm of pesticide penetration, propelling a paradigm shift in the understanding of food safety. Meanwhile, this strategy provides strong support for the cutting-edge application of SERS imaging technology and also brings valuable reference and enlightenment for researchers in related fields.

Visual whole-process monitoring of pesticide residues: An environmental perspective using surface-enhanced Raman spectroscopy with dynamic borohydride-reduced silver nanoparticles.

Environmental monitoring of pesticide residues in crops is essential for both food safety and environmental protection. Traditional methodologies face challenges due to the interference of endogenous compounds in peel and pulp tissues, often being invasive, labor-intensive, and inadequate for real-time observation of hazardous substance distribution. In this study, dynamic borohydride-reduced nanoparticles were employed as enhanced substrates. For the first time, surface-enhanced Raman spectroscopy (SERS) imaging was harnessed to enable whole-process visual detection of pesticide residues. The developed method is both stable and sensitive, boasting a detection lower limit below 1 pg/mL, coupled with robust quantitative analytical capabilities. This technique was successfully employed to detect residue signals across various crops and fruit juices. Furthermore, SERS imaging was utilized to map the distribution of pesticide residues from the exterior to the interior of fruits and vegetables. Vertex component analysis further refined the process by mitigating interference from plant autofluorescence. Collectively, this innovative strategy facilitates comprehensive pesticide residue monitoring, offering a potent tool for controlling hazardous substances in crops. Its potential applications extend beyond food safety, holding significant promise for sustainable agricultural production and enhanced environmental safeguarding.

Self-Modulated Ghost Imaging in Dynamic Scattering Media.

In this paper, self-modulated ghost imaging (SMGI) in a surrounded scattering medium is proposed. Different from traditional ghost imaging, SMGI can take advantage of the dynamic scattering medium that originally affects the imaging quality and generate pseudo-thermal light through the dynamic scattering of free particles' Brownian motion in the scattering environment for imaging. Theoretical analysis and simulation were used to establish the relationship between imaging quality and particle concentration. An experimental setup was also built to verify the feasibility of the SMGI. Compared with the reconstructed image quality and evaluation indexes of traditional ghost imaging, SMGI has better image quality, which demonstrates a promising future in dynamic high-scattering media such as dense fog and turbid water.

Microlens-Assisted Light-Scattering Imaging of Plasmonic Nanoparticles at the Single Particle Level.

We present a microlens-assisted imaging approach to record the scattering light of plasmonic nanoparticles at the single particle level. The microlens can significantly enhance the backscattering of visible light from individual plasmonic nanoparticles by several dozen folds, and single gold nanoparticles with a diameter as low as 60 nm can be imaged under a conventional optical microscope. This can benefit from a significant increase in the scattering intensity afforded by the microlens, meaning that the imaging of gold nanoparticles at a high temporal resolution (up to 5000 Hz) can be achieved, which is fast enough to record single particle adhesion events on the substrate. This research presents a fast and efficient means of acquiring scattering light from plasmonic nanoparticles, which has great potential to develop plasmonic nanoparticle-based biosensors and investigate a wide range of plasmonic nanoparticle-based fast interaction processes.

Analyzing the influence of oblique incidence on quantitative backscattering tissue polarimetry: a pilot ex vivo study.

Significance: Among the available polarimetric techniques, backscattering Mueller matrix (MM) polarimetry provides a promising non-contact and quantitative tool for in vivo tissue detection and clinical diagnosis. To eliminate the surface reflection from the sample cost-effectively, the non-collinear backscattering MM imaging setup always has an oblique incidence. Meanwhile, for practical organ cavities imaged using polarimetric gastrointestinal endoscopy, the uneven tissue surfaces can induce various relative oblique incidences inevitably, which can affect the polarimetry in a complicated manner and needs to be considered for detailed study. Aim: The purpose of this study is to systematically analyze the influence of oblique incidence on backscattering tissue polarimetry. Approach: We measured the MMs of experimental phantom and ex vivo tissues with different incident angles and adopted a Monte Carlo simulation program based on cylindrical scattering model for further verification and analysis. Meanwhile, the results were quantitatively evaluated using the Fourier transform, basic statistics, and frequency distribution histograms. Results: Oblique incidence can induce different changes on non-periodic, two-periodic, and four-periodic MM elements, leading to false-positive and false-negative polarization information for tissue polarimetry. Moreover, a prominent oblique incidence can bring more dramatic signal variations, such as phase retardance and element transposition. Conclusions: The findings presented in this study give some crucial criterions of appropriate incident angle selections for in vivo polarimetric endoscopy and other applications and can also be valuable references for studying how to minimize the influence further.

Plasmonic scattering imaging of single Cu2-xSe nanoparticle for Hg2+ detection.

The development of new efficient contrast nanoprobe has been greatly concerned in the field of scattering imaging for sensitive and accurate detection of trace analytes. In this work, the non-stoichiometric Cu2-xSe nanoparticle with typical localized surface plasmon resonance (LSPR) properties originating from their copper deficiency as a plasmonic scattering imaging probe was developed for sensitive and selective detection of Hg2+ under dark-field microscopy. Hg2+ can compete with Cu(I)/Cu(II) which were sources of optically active holes coexisting in these Cu2-xSe nanoparticles for its higher affinity with Se2-. The plasmonic properties of Cu2-xSe were adjusted effectively. Thus, the color scattering images of Cu2-xSe nanoparticles was changed from blue to cyan, and the scattering intensity was obviously enhanced with the dark-field microscopy. There was a linear relationship between the scattering intensity enhancement and the Hg2+ concentration in the range of 10-300 nM with a low detection limit of 1.07 nM. The proposed method has good potential for Hg2+ detection in the actual water samples. This work provides a new perspective on applying new plasmonic imaging probe for the reliable determination of trace heavy metal substances in the environment at a single particle level.

Microscopic Cascading Devices for Boosting Mucus Penetration in Oral Drug Delivery-Micromotors Nesting Inside Microcontainers.

In the case of macromolecules and poorly permeable drugs, oral drug delivery features low bioavailability and low absorption across the intestinal wall. Intestinal absorption can be improved if the drug formulation could be transported close to the epithelium. To achieve this, a cascade delivery device comprising Magnesium-based Janus micromotors (MMs) nesting inside a microscale containers (MCs) has been conceptualized. The device aims at facilitating targeted drug delivery mediated by MMs that can lodge inside the intestinal mucosa. Loading MMs into MCs can potentially enhance drug absorption through increased proximity and unidirectional release. The MMs will be provided with optimal conditions for ejection into any residual mucus layer that the MCs have not penetrated. MMS confined inside MCs propel faster in the mucus environment as compared to non-confined MMs. Upon contact with a suitable fuel, the MM-loaded MC itself can also move. An in vitro study shows fast release profiles and linear motion properties in porcine intestinal mucus compared to more complex motion in aqueous media. The concept of dual-acting cascade devices holds great potential in applications where proximity to epithelium and deep mucus penetration are needed.

Rapid Detection of Urinary Tract Infection in 10 min by Tracking Multiple Phenotypic Features in a 30 s Large-Volume Scattering Video of Urine Microscopy.

Rapid point-of-care (POC) diagnosis of bacterial infection diseases provides clinical benefits of prompt initiation of antimicrobial therapy and reduction of the overuse/misuse of unnecessary antibiotics for nonbacterial infections. We present here a POC compatible method for rapid bacterial infection detection in 10 min. We use a large-volume solution scattering imaging (LVSi) system with low magnifications (1-2×) to visualize bacteria in clinical samples, thus eliminating the need for culture-based isolation and enrichment. We tracked multiple intrinsic phenotypic features of individual cells in a short video. By clustering these features with a simple machine learning algorithm, we can differentiate Escherichia coli from similar-sized polystyrene beads, distinguish bacteria with different shapes, and distinguish E. coli from urine particles. We applied the method to detect urinary tract infections in 104 patient urine samples with a 30 s LVSi video, and the results showed 92.3% accuracy compared with the clinical culture results. This technology provides opportunities for rapid bacterial infection diagnosis at POC settings.

Investigation of image plane for image reconstruction of objects through diffusers via deep learning.

SIGNIFICANCE: The imaging of objects hidden in light-scattering media is a vital practical task in a wide range of applications, including biological imaging. Deep-learning-based methods have been used to reconstruct images behind scattering media under complex scattering conditions, but improvements in the quality of the reconstructed images are required. AIM: To investigate the effect of image plane on the accuracy of reconstructed images. APPROACH: Light reflected from an object passing through glass diffusers is captured by changing the image plane of an optical imaging system. Images are reconstructed by deep learning, and evaluated in terms of structural similarity index measure, classification accuracy of digital images, and training and testing error curves. RESULTS: The reconstruction accuracy was improved for the case in which the diffuser was imaged, compared to the case where the object was imaged. The training and testing error curves show that the loss converged to lower values in fewer epochs when the diffuser was imaged. CONCLUSIONS: The proposed approach demonstrates an improvement in the accuracy of the reconstruction of objects hidden through glass diffusers by imaging glass diffuser surfaces, and can be applied to objects at unknown locations in a scattering medium.

Intraoperative Assessment and Photothermal Ablation of the Tumor Margins Using Gold Nanoparticles.

Surgical resection is commonly used for therapeutic management of different solid tumors and is regarded as a primary standard of care procedure, but precise localization of tumor margins is a major intraoperative challenge. Herein, a generalized method by optimizing gold nanoparticles for intraoperative detection and photothermal ablation of tumor margins is introduced. These nanoparticles are detectable by highly sensitive surface-enhanced Raman scattering imaging. This non-invasive technique assists in delineating the two surgically challenged tumors in live mice with orthotopic colon or ovarian tumors. Any remaining residual tumors are also ablated by using post-surgical adjuvant photothermaltherapy (aPTT), which results in microscale heat generation due to interaction of these nanoparticles with near-infrared laser. Ablation of these post-operative residual micro-tumors prolongs the survival of mice significantly and delays tumor recurrence by 15 days. To validate clinical translatability of this method, the pharmacokinetics, biodistribution, Raman contrast, aPTT efficiency, and toxicity of these nanoparticles are also investigated. The nanoparticles have long blood circulation time (≈24 h), high tumor accumulation (4.87 ± 1.73%ID g-1) and no toxicity. This high-resolution and sensitive intraoperative approach is versatile and can be potentially used for targeted ablation of residual tumor after resection within different organs.

Quantification of Single-Molecule Protein Binding Kinetics in Complex Media with Prism-Coupled Plasmonic Scattering Imaging.

Measuring molecular binding is critical for understanding molecular-scale biological processes and screening drugs. Label-free detection technologies, such as surface plasmon resonance (SPR), have been developed for analyzing analytes in their natural forms. However, the specificity of these methods is solely relying on surface chemistry and has often nonspecific binding issues when working with samples in complex media. Herein, we show that single-molecule-based measurement can distinct specific and nonspecific binding processes by quantifying the mass and binding dynamics of individual-bound analyte molecules, thus allowing the binding kinetic analysis in complex media such as serum. In addition, this single-molecule imaging is realized in a commonly used Kretschmann prism-coupled SPR system, thus providing a convenient solution to realize high-resolution imaging on widely used prism-coupled SPR systems.

Advances in stimulated Raman scattering imaging for tissues and animals.

Stimulated Raman scattering (SRS) microscopy has emerged in the last decade as a powerful optical imaging technology with high chemical selectivity, speed, and subcellular resolution. Since the invention of SRS microscopy, it has been extensively employed in life science to study composition, structure, metabolism, development, and disease in biological systems. Applications of SRS in research and the clinic have generated new insights in many fields including neurobiology, tumor biology, developmental biology, metabolomics, pharmacokinetics, and more. Herein we review the advances and applications of SRS microscopy imaging in tissues and animals, as well as envision future applications and development of SRS imaging in life science and medicine.

Orientation-independent reaction activity monitoring with single particle and data analytics.

Single-particle analysis is the most powerful method to obtain accurate local information for understanding and monitoring chemical reactions. However, investigations about obtaining comprehensive information at the single-particle level to overcome individual errors and sampling randomness have not been reported to date. Plasmonic nanorods, which have excellent anisotropic optical and chemical properties, make us in situ acquisition of conformation and dynamics of the biological information. On the basis of their anisotropic optical properties of the plasmonic nanorods such as Au nanorods (AuNRs) and data analytics, herein we developed a high-throughput resonance scattering imaging method of AuNRs under dark-field microscopy (DFM) to monitor orientation-independent reaction activity of AuNRs. Data analytics are introduced to determine a large number of AuNRs orientation obtained from a series of polarized DFM images, allowing us to real-time monitor reaction activity of AuNRs at all orientations, and also makes it possible to study the global and local reaction processes of AuNRs at single-particle level. Our method is expected to provide a new strategy for analytical study and single-particle sensing in chemistry.

Calibration to Mitigate Near-Field Antennas Effects for a MIMO Radar Imaging System.

A calibration method for a high-resolution hybrid MIMO turntable radar imaging system is presented. A line of small metal spheres is employed as a test pattern in the calibration process to measure the position shift caused by undesired antenna effects. The unwanted effects in the antenna near-field responses are analysed, modelled and significantly mitigated based on the symmetry and differences in the responses of the MIMO configuration.

Nondestructive Detection for Egg Freshness Based on Hyperspectral Scattering Image Combined with Ensemble Learning.

Scattering hyperspectral technology is a nondestructive testing method with many advantages. Here, we propose a method to improve the accuracy of egg freshness, research the influence of incident angles of light source on the accuracy, and explain its mechanism. A variety of weak classifiers classify eggs based on the spectra after preprocessing and feature wavelength extraction to obtain three classifiers with the highest accuracy. The three classifiers are used as metamodels of stacking ensemble learning to improve the highest accuracy from 96.25% to 100%. Moreover, the highest accuracy of scattering, reflection, transmission, and mixed hyperspectral of eggs are 100.00%, 88.75%, 95.00%, and 96.25%, respectively, indicating that the scattering hyperspectral for egg freshness detection is better than that of the others. In addition, the accuracy is inversely proportional to the angle of incidence, i.e., the smaller the incident angle, the camera collects a larger proportion of scattering light, which contains more biochemical parameters of an egg than that of reflection and transmission. These results are very important for improving the accuracy of non-destructive testing and for selecting the incident angle of a light source, and they have potential applications for online non-destructive testing.

Dopamine-Mediated Assembly of Citrate-Capped Plasmonic Nanoparticles into Stable Core-Shell Nanoworms for Intracellular Applications.

Plasmonic nanochains, derived from the one-dimensional assembly of individual plasmonic nanoparticles (NPs), remain infrequently explored in biological investigations due to their limited colloidal stability, ineffective cellular uptake, and susceptibility to intracellular disassembly. We report the synthesis of polydopamine (PDA)-coated plasmonic "nanoworms" (NWs) by sonicating citrate-capped gold (Cit-Au) NPs in a concentrated dopamine (DA) solution under alkaline conditions. DA mediates the assembly of Cit-Au NPs into Au NWs within 1 min, and subsequent self-polymerization of DA for 60 min enables the growth of an outer conformal PDA shell that imparts stability to the inner Au NW structure in solution, yielding "core-shell" Au@PDA NWs with predominantly 4-5 Au cores per worm. Our method supports the preparation of monometallic Au@PDA NWs with different core sizes and bimetallic PDA-coated NWs with Au and silver cores. The protonated primary amine and catechol groups of DA, with their ability to interact with Cit anions via hydrogen bonding and electrostatic attraction, are critical to assembly. When compared to unassembled PDA-coated Au NPs, our Au@PDA NWs scatter visible light and absorb near-infrared light more intensely and enter HeLa cancer cells more abundantly. Au@PDA NWs cross the cell membrane as intact entities primarily via macropinocytosis, mostly retain their inner NW structure and outer PDA shell inside the cell for 24 h, and do not induce noticeable cytotoxicity. We showcase three intracellular applications of Au@PDA NWs, including label-free dark-field scattering cell imaging, delivery of water-insoluble cargos without pronounced localization in acidic compartments, and photothermal killing of cancer cells.

Facile synthesis of terminal-alkyne bioorthogonal molecules for live -cell surface-enhanced Raman scattering imaging through Au-core and silver/dopamine-shell nanotags.

Alkyne is unique, specific and biocompatible in the Raman-silent region of the cell, but there still remains a challenge to achieve ultrasensitive detection in living systems due to its weak Raman scattering. Herein, a terminal alkyne ((E)-2-[4-(ethynylbenzylidene)amino]ethane-1-thiol (EBAE)) with surface-enhanced Raman scattering is synthesized. The EBAE molecule possesses S- and C-termini, which can be directly bonded to gold nanoparticles and dopamine/silver by forming the Au-S chemical bond and the carbon-metal bond, respectively. The distance between Raman reporter and AuNPs/AgNPs can be reduced, contributing to forming hot-spot-based SERS substrate. The alkyne functionalized nanoparticles are based on Au core and encapsulating polydopamine shell, defined as Au-core and dopamine/Ag-shell (ACDS). The bimetallic ACDS induce strong SERS signals for molecular imaging that arise from the strong electromagnetic field. Furthermore, the EBAE provides a distinct peak in the cellular Raman-silent region with nearly zero background interference. The EBAE Raman signals could be tremendously enhanced when the Raman reporter is located at the middle of the Au-core and dopamine/Ag-shell. Therefore, this work could have huge potential benefits for the highly sensitive detection of intercellular information delivery by connecting the recognition molecules in biomedical diagnostics. Graphical abstract Terminal-alkyne-functionalized Au-core and silver/dopamine-shell nanotags for live-cell surface-enhanced Raman scattering imaging.

A rapid and simple method for efficient capture and accurate discrimination of circulating tumor cells using aptamer conjugated magnetic beads and surface-enhanced Raman scattering imaging.

The efficient isolation and the accurate phenotype discrimination of circulating tumor cells (CTCs) are expected to provide much valuable information for the understanding of tumor metastasis and to play an important role in personalized treatment of cancer patients in the future. In this study, we developed a novel, rapid, and simple method for efficient capture and accurate identification of CTCs using aptamer conjugated magnetic beads and surface-enhanced Raman scattering (SERS) imaging technique. Using aptamer conjugated magnetic beads, rare target cancer cells can be captured efficiently from buffer and whole blood sample with capture efficiency of 73 % and 55 %, respectively. Meanwhile, captured cancer cells were labeled by specific SERS probes and can be identified readily and accurately by SERS imaging technique. Results of our experiment demonstrate the potential feasibility of aptamer conjugated magnetic beads coupled with SERS imaging technique for the efficient capture and accurate discrimination of CTCs in clinical whole blood sample. Graphical Abstract Schematic Representation of CTCs Capture and Identification Using Apt-MBs and SERS Imaging.

In situ micro-spectroscopic investigation of lignin in poplar cell walls pretreated by maleic acid.

BACKGROUND: In higher plant cells, lignin provides necessary physical support for plant growth and resistance to attack by microorganisms. For the same reason, lignin is considered to be a major impediment to the process of deconstructing biomass to simple sugars by hydrolytic enzymes. The in situ variation of lignin in plant cell walls is important for better understanding of the roles lignin play in biomass recalcitrance. RESULTS: A micro-spectroscopic approach combining stimulated Raman scattering microscopy and fluorescence lifetime imaging microscopy was employed to probe the physiochemical structure of lignin in poplar tracheid cell walls. Two forms of lignins were identified: loosely packed lignin, which had a long (4 ns) fluorescence lifetime and existed primarily in the secondary wall layers; and dense lignin, which had a short (0.5-1 ns) fluorescence lifetime and was present in all wall layers, including the cell corners, compound middle lamellae, and secondary wall. At low maleic acid concentration (0.025 and 0.05 M) pretreatment conditions, some of the dense lignin was modified to become more loosely packed. High acid concentration removed both dense and loosely packed lignins. These modified lignins reformed to make lignin-carbohydrate complex droplets containing either dense or loosely packed lignin (mostly from secondary walls) and were commonly observed on the cell wall surface. CONCLUSIONS: We have identified dense and loosely packed lignins in plant cell walls. During maleic acid pretreatment, both dense lignin droplets and loosely packed lignin droplets were formed. Maleic acid pretreatment more effectively removes loosely packed lignin in secondary walls which increases enzyme accessibility for digestion.