From Self-Assembly of Colloidal Crystals toward Ordered Porous Layer Interferometry.

Interferometry-based, reflectometric, label-free biosensors have made significant progress in the analysis of molecular interactions after years of development. The design of interference substrates is a key research topic for these biosensors, and many studies have focused on porous films prepared by top-down methods such as porous silicon and anodic aluminum oxide. Lately, more research has been conducted on ordered porous layer interferometry (OPLI), which uses ordered porous colloidal crystal films as interference substrates. These films are made using self-assembly techniques, which is the bottom-up approach. They also offer several advantages for biosensing applications, such as budget cost, adjustable porosity, and high structural consistency. This review will briefly explain the fundamental components of self-assembled materials and thoroughly discuss various self-assembly techniques in depth. We will also summarize the latest studies that used the OPLI technique for label-free biosensing applications and divide them into several aspects for further discussion. Then, we will comprehensively evaluate the strengths and weaknesses of self-assembly techniques and discuss possible future research directions. Finally, we will outlook the upcoming challenges and opportunities for label-free biosensing using the OPLI technique.

Drug screening at single-organoid resolution via bioprinting and interferometry.

High throughput drug screening is an established approach to investigate tumor biology and identify therapeutic leads. Traditional platforms use two-dimensional cultures which do not accurately reflect the biology of human tumors. More clinically relevant model systems such as three-dimensional tumor organoids can be difficult to scale and screen. Manually seeded organoids coupled to destructive endpoint assays allow for the characterization of treatment response, but do not capture transitory changes and intra-sample heterogeneity underlying clinically observed resistance to therapy. We present a pipeline to generate bioprinted tumor organoids linked to label-free, time-resolved imaging via high-speed live cell interferometry (HSLCI) and machine learning-based quantitation of individual organoids. Bioprinting cells gives rise to 3D structures with unaltered tumor histology and gene expression profiles. HSLCI imaging in tandem with machine learning-based segmentation and classification tools enables accurate, label-free parallel mass measurements for thousands of organoids. We demonstrate that this strategy identifies organoids transiently or persistently sensitive or resistant to specific therapies, information that could be used to guide rapid therapy selection.

Ultrasensitive deafness gene DNA hybridization detection employing a fiber optic Mach-Zehnder interferometer: Enabled by a black phosphorus nanointerface.

The rapid and efficient detection of deafness gene DNA plays an important role in the clinical diagnosis of deafness diseases. This study demonstrates the ultrasensitive detection of complementary DNA (cDNA) by employing a nanointerface-sensitized fiber optic biosensor. The sensor consists of SMF-TNCF-MMF-SMF (abbreviated as STMS) structure with lateral offset. Besides, it is functionalized with a nanointerface of black phosphorus (BP) to enhance the light-matter interaction and eventually improve the sensing performances. Relying on this nanointerface-sensitized sensor, we successfully realize the in-situ detection of cDNA at concentrations ranging from 1 pM to 1 µM, with a sensitivity of 0.719 nm/lgM. The limit of detection (LOD) is as low as 0.24 pM, which is at least two orders of magnitude lower than those of existing methods. The sensor exhibits the advantages of simple operation, fast response, label-free measurement, excellent repeatability, and high selectivity. Our contribution suggests a convenient approach for deafness gene DNA detection and can be extended for general ultra-low concentration DNA detection applications.

Lava Mapping Using Sentinel-1 Data after the Occurrence of a Volcanic Eruption-The Case of Cumbre Vieja Eruption on La Palma, Canary Islands, Spain.

Volcanic eruptions pose a great threat to humans. In this context, volcanic hazard and risk assessment constitute crucial issues with respect to mitigating the effects of volcanic activity and ensuring the health and safety of inhabitants. Lava flows directly affect communities living near active volcanoes. Nowadays, remote sensing advances make it possible to effectively monitor eruptive activity, providing immediate and accurate information concerning lava evolution. The current research focuses on the mapping of the surface deformation and the analysis of lava flow evolution occurred on the island of La Palma, during the recent (2021) eruptive phase of the volcano. Sentinel-1 data covering the island were collected throughout the entire eruptive period, i.e., September 2021 until January 2022. The processing was based on amplitude-based and phase-based detection methods, i.e., Synthetic Aperture Radar interferometry (InSAR) and offset tracking. In particular, ground deformation occurred on the island, while Line-Of-Sight (LOS) displacements were derived from Sentinel-1 interferograms. Moreover, the evolution of lava flow velocity was estimated using Sentinel-1 imagery along with offset tracking technique. The maximum lava flow velocity was calculated to be 2 m/day. It was proved that both approaches can provide rapid and useful information in emergencies, especially in inaccessible areas. Although offset tracking seems a quite promising technique for the mapping of lava flows, it still requires improvement.

Real-time monitoring of the hydrolysis of vegetable oils loaded in silica colloidal crystal films with lipase by optical interferometry.

Different vegetable oils have different nutritional components, especially in terms of the composition of their fatty acids, which can only be reflected after entering the human body. Therefore, when judging their health value and identifying high-quality vegetable oils, in addition to the analysis of their ingredients, tracking their hydrolysis process in the human body is a very important aspect. However, most identification methods or simulated digestion studies fail to achieve this. In this paper, we applied ordered porous layer interferometry (OPLI) for the real-time monitoring of optical thickness changes (ΔOT) to track the hydrolysis process of four kinds of vegetable oil. Further, this study can obtain precise data (the initial rate and degree of hydrolysis) to provide more information on the hydrolysis ability of different vegetable oils and give references for their nutritional and functional evaluation. In addition, it provides more possibilities for the adulteration identification and bioavailability analysis of vegetable oils.

Real-time monitoring of immunoglobulin G levels in milk using an ordered porous layer interferometric optical sensor.

Immunoglobulin G (IgG) is a significant ingredient of immunological activity in milk and colostrum, the activity and content of which is easily disturbed by potentially conditional variant during sterilization. Therefore, developing robust methods for the detection of IgG levels in milk is especially important. Herein, protein A from the Staphylococcus aureus functionalized silica colloidal crystalline film (SCC@SPA) sensing unit combined with ordered porous layer interferometry (OPLI) for IgG detection in untreated bovine milk was developed. Calibration curves in milk and buffer were set up by the variations of the optical thickness (OT) of the sensing unit after the IgG association and dissociation phases. The influence of temperature on the level of IgG was evaluated. Furthermore, the identification of IgG levels with pasteurized milk and ultrahigh temperature (UHT) sterilized milk from the market randomly was successfully carried out without any sample pretreatment. More importantly, compared with other methods, this novel method has the advantages of convenient operation, low cost, and suitability for point-of-care (POC) testing.

Simultaneous directional full-field OCT using path-length and carrier multiplexing.

Full-field swept-source optical coherence tomography (FF-SS-OCT) is an emerging technology with potential applications in ophthalmic imaging, microscopy, metrology, and other domains. Here we demonstrate a novel method of multiplexing FF-SS-OCT signals using carrier modulation (CM). The principle of CM could be used to inspect various properties of the scattered light, e.g. its spectrum, polarization, Doppler shift, or distribution in the pupil. The last of these will be explored in this work, where CM was used to acquire images passing through two different optical pupils. The two pupils contained semicircular optical windows with perpendicular orientations, with each window permitting measurement of scattering anisotropy in one dimension by inducing an optical delay between the images formed by the two halves of the pupil. Together, the two forms of multiplexing permit measurement of differential scattering anisotropy in the x and y dimensions simultaneously. To demonstrate the feasibility of this technique our carrier multiplexed directional FF-OCT (CM-D-FF-OCT) system was used to acquire images of a microlens array, human hair, onion skin and in vivo human retina. The results of these studies are presented and briefly discussed in the context of future development and application of this technique.

Identification of natural compounds as SARS-CoV-2 entry inhibitors by molecular docking-based virtual screening with bio-layer interferometry.

Coronavirus Disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which enter the host cells through the interaction between its receptor binding domain (RBD) of spike glycoprotein with angiotensin-converting enzyme 2 (ACE2) receptor on the plasma membrane of host cell. Neutralizing antibodies and peptide binders of RBD can block viral infection, however, the concern of accessibility and affordability of viral infection inhibitors has been raised. Here, we report the identification of natural compounds as potential SARS-CoV-2 entry inhibitors using the molecular docking-based virtual screening coupled with bilayer interferometry (BLI). From a library of 1871 natural compounds, epigallocatechin gallate (EGCG), 20(R)-ginsenoside Rg3 (RRg3), 20(S)-ginsenoside Rg3 (SRg3), isobavachalcone (Ibvc), isochlorogenic A (IscA) and bakuchiol (Bkc) effectively inhibited pseudovirus entry at concentrations up to 100 µM. Among these compounds, four compounds, EGCG, Ibvc, salvianolic acid A (SalA), and isoliensinine (Isl), were effective in inhibiting SARS-CoV-2-induced cytopathic effect and plaque formation in Vero E6 cells. The EGCG was further validated with no observable animal toxicity and certain antiviral effect against SARS-CoV-2 pseudovirus mutants (D614G, N501Y, N439K & Y453F). Interestingly, EGCG, Bkc and Ibvc bind to ACE2 receptor in BLI assay, suggesting a dual binding to RBD and ACE2. Current findings shed some insight into identifications and validations of SARS-CoV-2 entry inhibitors from natural compounds.

A Miniature Bio-Photonics Companion Diagnostics Platform for Reliable Cancer Treatment Monitoring in Blood Fluids.

In this paper, we present the development of a photonic biosensor device for cancer treatment monitoring as a complementary diagnostics tool. The proposed device combines multidisciplinary concepts from the photonic, nano-biochemical, micro-fluidic and reader/packaging platforms aiming to overcome limitations related to detection reliability, sensitivity, specificity, compactness and cost issues. The photonic sensor is based on an array of six asymmetric Mach Zender Interferometer (aMZI) waveguides on silicon nitride substrates and the sensing is performed by measuring the phase shift of the output signal, caused by the binding of the analyte on the functionalized aMZI surface. According to the morphological design of the waveguides, an improved sensitivity is achieved in comparison to the current technologies (<5000 nm/RIU). This platform is combined with a novel biofunctionalization methodology that involves material-selective surface chemistries and the high-resolution laser printing of biomaterials resulting in the development of an integrated photonics biosensor device that employs disposable microfluidics cartridges. The device is tested with cancer patient blood serum samples. The detection of periostin (POSTN) and transforming growth factor beta-induced protein (TGFBI), two circulating biomarkers overexpressed by cancer stem cells, is achieved in cancer patient serum with the use of the device.

Effects of Vectored Thermal Pulsation on Objective Tear Film Measures.

PURPOSE: To assess the effect of vectored thermal pulsation treatment for meibomian gland dysfunction on objective measures of lipid layer thickness (LLT) and tear osmolarity. METHODS: One hundred patients with meibomian gland dysfunction diagnosis were recruited to participate. At their initial visit, baseline study parameters were recorded, and vectored thermal pulsation treatment was administered. At the 2- to 3-month follow-up visit, the study parameters were reevaluated. Subjective symptoms were evaluated using the Ocular Surface Disease Index questionnaire. LLT was measured using an ocular surface interferometer. Tear osmolarity was calculated using impedance measurement of tear fluid collected from the eyelid margin. RESULTS: Ninety-six patients (192 eyes) completed the follow-up. Mean improvement in Ocular Surface Disease Index was 5.6 points (95% confidence interval [CI], -9.0 to -2.1, P = 0.002). There was no significant change in tear osmolarity (mean change -1.6 mOsm/L, 95% CI, -4.7 to +1.3 mOsm/L, P = 0.3). There was no significant change in LLT (mean change -4.3 nm, 95% CI, -9.1 to +0.5 nm, P = 0.08). CONCLUSIONS: The hypothesis that vectored thermal pulsation treatment would decrease tear osmolarity and increase LLT was not substantiated. Although we detected significant improvement in subjective symptoms, the improvement was smaller than the improvements reported in previous studies. Our results suggest that the current understanding of the effects of vectored thermal pulsation treatment is incomplete.

Empirical beam hardening and ring artifact correction for x-ray grating interferometry (EBHC-GI).

PURPOSE: Talbot-Lau grating interferometry enables the use of polychromatic x-ray sources, extending the range of potential applications amenable to phase contrast imaging. However, these sources introduce beam hardening effects not only from the samples but also from the gratings. As a result, grating inhomogeneities due to manufacturing imperfections can cause spectral nonuniformity artifacts when used with polychromatic sources. Consequently, the different energy dependencies of absorption, phase, and visibility contrasts impose challenges that so far have limited the achievable image quality. The purpose of this work was to develop and validate a correction strategy for grating-based x-ray imaging that accounts for beam hardening generated from both the imaged object and the gratings. METHODS: The proposed two-variable polynomial expansion strategy was inspired by work performed to address beam hardening from a primary modulator. To account for the multicontrast nature of grating interferometry, this approach was extended to each contrast to obtain three sets of correction coefficients, which were determined empirically from a calibration scan. The method's feasibility was demonstrated using a tabletop Talbot-Lau grating interferometer micro-computed tomography (CT) system using CT acquisitions of a water sample and a silicon sample, representing low and high atomic number materials. Spectral artifacts such as cupping and ring artifacts were quantified using mean squared error (MSE) from the beam-hardening-free target image and standard deviation within a reconstructed image of the sample. Finally, the model developed using the water sample was applied to a fixated murine lung sample to demonstrate robustness for similar materials. RESULTS: The water sample's absorption CT image was most impacted by spectral artifacts, but following correction to decrease ring artifacts, an 80% reduction in MSE and 57% reduction in standard deviation was observed. The silicon sample created severe artifacts in all contrasts, but following correction, MSE was reduced by 94% in absorption, 96% in phase, and 90% in visibility images. These improvements were due to the removal of ring artifacts for all contrasts and reduced cupping in absorption and phase images and reduced capping in visibility images. When the water calibration coefficients were applied to the lung sample, ring artifacts most prominent in the absorption contrast were eliminated. CONCLUSIONS: The described method, which was developed to remove artifacts in absorption, phase, and normalized visibility micro-CT images due to beam hardening in the system gratings and imaged object, reduced the MSE by up to 96%. The method depends on calibrations that can be performed on any system and does not require detailed knowledge of the x-ray spectrum, detector energy response, grating attenuation properties and imperfections, or the geometry and composition of the imaged object.

Time-of-flight resolved light field fluctuations reveal deep human tissue physiology.

Red blood cells (RBCs) transport oxygen to tissues and remove carbon dioxide. Diffuse optical flowmetry (DOF) assesses deep tissue RBC dynamics by measuring coherent fluctuations of multiply scattered near-infrared light intensity. While classical DOF measurements empirically correlate with blood flow, they remain far-removed from light scattering physics and difficult to interpret in layered media. To advance DOF measurements closer to the physics, here we introduce an interferometric technique, surmounting challenges of bulk motion to apply it in awake humans. We reveal two measurement dimensions: optical phase, and time-of-flight (TOF), the latter with 22 picosecond resolution. With this multidimensional data, we directly confirm the unordered, or Brownian, nature of optically probed RBC dynamics typically assumed in classical DOF. We illustrate how incorrect absorption assumptions, anisotropic RBC scattering, and layered tissues may confound classical DOF. By comparison, our direct method enables accurate and comprehensive assessment of blood flow dynamics in humans.

Highly sensitive IRS based biosensor for the determination of cytochrome c as a cancer marker by using nanoporous anodic alumina modified with trypsin.

The determination of cytochrome c in the human serum sample is a regular medical investigation performed to assess cancer diseases. Herein, we used interferometric reflectance spectroscopy (IRS) based biosensor for the determination of cytochrome c. For this purpose first, the nanoporous anodic alumina (NAA) was fabricated. Then, the NAA pore walls were functionalized with 3-aminopropyl trimethoxy silane (NAA-NH2). Subsequently, the trypsin enzyme was immobilized on the NAA pore walls. The sensing principle of proposed IRS sensor to cytochrome c is based on a change in the intensity of the reflected light to a charge-coupled device (CCD) detector after digesting of cytochrome c by immobilized trypsin enzymes on NAA-NH2 into the heme-peptide fragment. The heme-peptide fragment then oxidized 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) to green color ABTS·- anion radical in the presence of hydrogen peroxide. The generated green color ABTS·- anion radical solution adsorbed the white light and therefore the intensity of the reflected light from NAA to the CCD decreased. The decrease in the intensity of the white light had a logarithmic relationship with the concentration of the cytochrome c in the range of 1-100 nM. The limit of detections (LOD) for cytochrome c was 0.5 nM. The proposed biosensor exhibited high selectivity, sensitivity, and good stability.

Aptamer-Based Nanoporous Anodic Alumina Interferometric Biosensor for Real-Time Thrombin Detection.

Aptamer biosensors are one of the most powerful techniques in biosensing. Achieving the best platform to use in aptamer biosensors typically includes crucial chemical modifications that enable aptamer immobilization on the surface in the most efficient manner. These chemical modifications must be well defined. In this work we propose nanoporous anodic alumina (NAA) chemically modified with streptavidin as a platform for aptamer immobilization. The immobilization of biotinylated thrombin binding aptamer (TBA) was monitored in real time by means of reflective interferometric spectroscopy (RIfS). The study has permitted to characterize in real time the path to immobilize TBA on the inner pore walls of NAA. Furthermore, this study provides an accurate label-free method to detect thrombin in real-time with high affinity and specificity.

Osteointegrative and microgeometric comparison between micro-blasted and alumina blasting/acid etching on grade II and V titanium alloys (Ti-6Al-4V).

This study evaluated the effect of alumina-blasted/acid-etched (AB/AE) or microabrasive blasting (C3-Microblasted) surface treatment on the osseointegration of commercially-pure Ti (grade II) and Ti-6Al-4V alloy (grade V) implants compared to as-machined surfaces. Surface characterization was performed by scanning electron microscopy and optical interferometry (IFM) to determine roughness parameters (Sa and Sq, n = 3 per group). One-hundred forty-four implants were placed in the radii of 12 beagle dogs, for histological (n = 72, bone-to-implant contact - BIC and bone-area-fraction occupancy -BAFO) and torque to interface failure test at 3 and 6 weeks (n = 72). SEM and IFM revealed a significant increase in surface texture for AB/AE and C3-Microblasted surfaces compared to machined surface, regardless of titanium substrate. Torque-to-interface failure test showed significant increase in values from as-machined to AB/AE and to C3-Microblasted. Considering time in vivo, alloy grade, and surface treatment, the C3-microblasted presented higher mean BIC values relative to AB/AE and machined surfaces for both alloy types. BAFO levels were significantly higher for both textured surfaces groups relative to the machined group at 3 weeks, but differences were not significant between the three surfaces for each alloy type at 6 weeks. Surface treatment resulted in roughness that improved osseointegration in Grade II and V titanium substrates.

Establishment of Logic Gates Based on Conformational Changes in a Multiple-Factor Biomolecule Interaction Process by Dual Polarization Interferometry.

DNA-based logic gates stimulate the development of molecular scale computers and show enormous potential in nanotechnology, biotechnology, and medicine. However, the reported detectors to date usually require one to label appropriate signal probes, resulting in not only a high cost but also potentially tedious manipulation. For the first time, we established a label-free logic gate by regarding the structure-related signal as output. Dual polarization interferometry (DPI) was employed to reveal the detailed conformational transitions occurring in the multiple-factor biomolecule interactions and then was utilized as a detection tool of logic gate. As a vital merit of this system, the dependence of the density output signal on the interaction with multiple-factor input can mimic the function of signal communication in OR, INHIBIT, and IDENTITY logic gates and the INHIBIT-OR cascade circuit. Additionally, the DPI signal with logic stringency can unambiguously distinguish conformational polymorphisms and compare structural stability. This study provides a new way for the construction of a label-free logic gate, supplements information deficiency of reaction details, and extends the application of DPI in logic operation.

Environmental Copper Sensor Based on Polyethylenimine-Functionalized Nanoporous Anodic Alumina Interferometers.

Anthropogenic copper pollution of environmental waters from sources such as acid mine drainage, antifouling paints, and industrial waste discharge is a major threat to our environment and human health. This study presents an optical sensing system that combines self-assembled glutaraldehyde-cross-linked double-layered polyethylenimine (PEI-GA-PEI)-modified nanoporous anodic alumina (NAA) interferometers with reflectometric interference spectroscopy (RIfS) for label-free, selective monitoring of ionic copper in environmental waters. Calibration of the sensing system with analytical solutions of copper shows a linear working range between 1 and 100 mg L-1, and a low limit of detection of 0.007 ± 0.001 mg L-1 (i.e., ∼0.007 ppm). Changes in the effective optical thickness (ΔOTeff) of PEI-GA-PEI-functionalized NAA interferometers are monitored in real-time by RIfS, and correlated with the amount of ionic copper present in aqueous solutions. The system performance is validated through X-ray photoelectron spectroscopy (XPS) and the spatial distribution of copper within the nanoporous films is characterized by time-of-flight-secondary ion mass spectroscopy (TOF-SIMS). The specificity and chemical selectivity of the PEI-GA-PEI-NAA sensor to Cu2+ ions is verified by screening six different metal ion solutions containing potentially interfering ions such as Al3+, Cd2+, Fe3+, Pb2+, Ni2+, and Zn2+. Finally, the performance of the PEI-GA-PEI-NAA sensor for real-life applications is demonstrated using legacy acid mine drainage liquid and tap water for qualitative and quantitative detection of copper ions. This study provides new opportunities to develop portable, cost-competitive, and ultrasensitive sensing systems for real-life environmental applications.

Label-free real-time optical monitoring of DNA hybridization using SiN Mach-Zehnder interferometer-based integrated biosensing platform.

We report on the label-free real-time optical monitoring of DNA hybridization upon exposure to a flow of complementary DNA at different concentrations. The biosensor is composed of a silicon nitride integrated unbalanced Mach-Zehnder interferometer (MZI), with an integrated arrayed waveguide grating as a spectral filter. This MZI has been shown to have both sufficient multiplexing capability and limit of detection on the order of 10 - 6 RIU. Probe DNA, consisting of a 36-mer fragment is covalently immobilized on the silicon nitride integrated biosensor. The wavelength shift is monitored upon complementary DNA targets being flown over the sensor. Concentrations of 1 pM can be easily detected. Also, an alternative route to modify the sensor surface with carboxylic groups using the photochemical reaction of fatty acids is proposed and preliminary XPS results are presented. Moreover, preliminary results for DNA obtained from a rolling circle amplification (RCA-DNA) process and spiked in a realistic amplification buffer are presented.

Silicon Photonic Biosensors Using Label-Free Detection.

Thanks to advanced semiconductor microfabrication technology, chip-scale integration and miniaturization of lab-on-a-chip components, silicon-based optical biosensors have made significant progress for the purpose of point-of-care diagnosis. In this review, we provide an overview of the state-of-the-art in evanescent field biosensing technologies including interferometer, microcavity, photonic crystal, and Bragg grating waveguide-based sensors. Their sensing mechanisms and sensor performances, as well as real biomarkers for label-free detection, are exhibited and compared. We also review the development of chip-level integration for lab-on-a-chip photonic sensing platforms, which consist of the optical sensing device, flow delivery system, optical input and readout equipment. At last, some advanced system-level complementary metal-oxide semiconductor (CMOS) chip packaging examples are presented, indicating the commercialization potential for the low cost, high yield, portable biosensing platform leveraging CMOS processes.

Combined high-speed holographic shape and full-field displacement measurements of tympanic membrane.

The conical shape of the tympanic membrane (TM or eardrum) plays an important role in its function, such that variations in shape alter the acoustically induced motions of the TM. We present a method that precisely determines both shape and acoustically induced transient response of the entire TM using the same optics and maintaining the same coordinate system, where the TM transient displacements due to a broadband acoustic click excitation (50-µs impulse) and the shape are consecutively measured within <200 ms. Interferograms gathered with continuous high-speed (>2 kHz) optical phase sampling during a single 100-ms wavelength tuning ramp allow precise and rapid reconstructions of the TM shape at varied resolutions (50 to 200 µm). This rapid acquisition of full-field displacements and shape is immune to slow disturbances introduced by breathing or heartbeat of live subjects. Knowledge of TM shape and displacements enables the estimation of surface normal displacements regardless of the orientation of the TM within the measurement system. The proposed method helps better define TM mechanics and provides TM structure and function information useful for the diagnosis of ear disease.