On-chip hybrid integration of swept frequency distributed-feedback laser with silicon photonic circuits using photonic wire bonding.

This paper presents a novel co-packaging approach through on-chip hybrid laser integration with photonic circuits using photonic wire bonding. The process involves die-bonding a low-cost semiconductor distributed-feedback (DFB) laser into a deep trench on a silicon-on-insulator (SOI) chip and coupling it to the silicon circuitry through photonic wire bonding (PWB). After characterizing the power-current-voltage (LIV) and optical spectrum of the laser, a wavelength-current relationship utilizing its tunability through self-heating a swept-frequency laser (SFL) is developed. Photonic integrated circuit (PIC) resonators are successfully characterized using the SFL method, demonstrating signal detection with a quality factor comparable to measurements conducted with an off-chip benchtop laser.

Best Practices for Germicidal Ultraviolet-C Dose Measurement for N95 Respirator Decontamination.

Ultraviolet-C (UV-C) decontamination holds promise in combating the coronavirus disease 2019 pandemic, particularly with its potential to mitigate the N95 respirator shortage. Safe, effective, and reproducible decontamination depends critically on UV-C dose, yet dose is frequently measured and reported incorrectly, which results in misleading and potentially harmful protocols. Understanding best practices in UV-C dose measurement for N95 respirator decontamination is essential to the safety of medical professionals, researchers, and the public. Here, we outline the fundamental optical principles governing UV-C irradiation and detection, as well as the key metrics of UV-C wavelength and dose. In particular, we discuss the technical and regulatory distinctions between UV-C N95 respirator decontamination and other applications of germicidal UV-C, and we highlight the unique considerations required for UV-C N95 respirator decontamination. Together, this discussion will inform best practices for UV-C dose measurement for N95 respirator decontamination during crisis-capacity conditions.

Design and fabrication of SOI micro-ring resonators based on sub-wavelength grating waveguides.

Standard silicon photonic strip waveguides offer a high intrinsic refractive index contrast; this permits strong light confinement, leading to compact bends, which in turn facilitates the fabrication of devices with small footprints. Sub-wavelength grating (SWG) based waveguides can allow the fabrication of low loss devices with specific, engineered optical properties. The combination of SWG waveguides with optical micro-resonators can offer the possibility of achieving resonators with properties different from the traditional SOI rings. One important property that SWG rings can offer is decreased light confinement in the waveguide core; this improves the resonator's sensitivity to changes in the cladding refractive index, making the rings ideal for refractive index sensing applications. In this paper, we present the design and experimental characterization of SWG based rings realized on SOI chips without upper cladding (permitting their use as sensors). The fabricated rings offer quality factors in the range of ~1k-6k, depending on SWG parameters. Based on the comparison of experimental and simulated data we expect sensitivities exceeding 383 nm/RIU in water and 270 nm/RIU in air, showing excellent potential for use in sensing applications.

Alginate core-shell beads for simplified three-dimensional tumor spheroid culture and drug screening.

We demonstrate that when using cell-laden core-shell hydrogel beads to support the generation of tumor spheroids, the shell structure reduces the out-of-bead and monolayer cell proliferation that occurs during long-term culture of tumor cells within core-only alginate beads. We fabricate core-shell beads in a two-step process using simple, one-layer microfluidic devices. Tumor cells encapsulated within the bead core will proliferate to form multicellular aggregates which can serve as three-dimensional (3-D) models of tumors in drug screening. Encapsulation in an alginate shell increased the time that cells could be maintained in three-dimensional culture for MCF-7 breast cancer cells prior to out-of-bead proliferation, permitting formation of spheroids over a period of 14 days without the need move the cell-laden beads to clean culture flasks to separate beads from underlying monolayers. Tamoxifen and docetaxel dose response shows decreased toxicity for multicellular aggregates in three-dimensional core-shell bead culture compared to monolayer. Using simple core-only beads gives mixed monolayer and 3-D culture during drug screening, and alters the treatment result compared to the 3-D core-shell or the 2-D monolayer groups, as measured by standard proliferation assay. By preventing the out-of-bead proliferation and subsequent monolayer formation that is observed with core-only beads, the core-shell structure can obviate the requirement to transfer the beads to a new culture flask during drug screening, an important consideration for cell-based drug screening and drugs which have high multicellular resistance index.

Designing a Microfluidic Device with Integrated Ratiometric Oxygen Sensors for the Long-Term Control and Monitoring of Chronic and Cyclic Hypoxia.

Control of oxygen over cell cultures in vitro is a topic of considerable interest, as chronic and cyclic hypoxia can alter cell behaviour. Both static and transient hypoxic levels have been found to affect tumour cell behaviour; it is potentially valuable to include these effects in early, in vitro stages of drug screening. A barrier to their inclusion is that rates of transient hypoxia can be a few cycles/hour, which is difficult to reproduce in traditional in vitro cell culture environments due to long diffusion distances from control gases to the cells. We use a gas-permeable three-layer microfluidic device to achieve spatial and temporal oxygen control with biologically-relevant switching times. We measure the oxygen profiles with integrated, ratiometric optical oxygen sensors, demonstrate sensor and system stability over multi-day experiments, and characterize a pre-bleaching process to improve sensor stability. We show, with both finite-element modelling and experimental data, excellent control over the oxygen levels by the device, independent of fluid flow rate and oxygenation for the operating flow regime. We measure equilibration times of approximately 10 min, generate complex, time-varying oxygen profiles, and study the effects of oxygenated media flow rates on the measured oxygen levels. This device could form a useful tool for future long-term studies of cell behaviour under hypoxia.

Sub-wavelength grating components for integrated optics applications on SOI chips.

In this paper we demonstrate silicon on insulator (SOI) sub-wavelength grating (SWG) optical components for integrated optics and sensing. Light propagation in SWG devices is studied and realized with no cladding on top of the waveguide. In particular, we focused on SWG bends, tapers and directional couplers, all realized with compatible geometries in order to be used as building blocks for more complex integrated optics devices (interferometers, switches, resonators, etc.). Fabricated SWG tapers for TE and TM polarizations are described; they allow for connecting SWG devices to regular strip waveguides with loss lower than 1 dB per taper. Our SWG directional coupler presents a very compact design and a negligible wavelength dependence of its crossover length (and as a consequence of its coupling coefficient, κ), over a 40 nm bandwidth. This wavelength flatten response represents a bandwidth enhancement with respect to standard directional couplers (made using strip or rib waveguides), in particular for the TE mode. SWG bends are demonstrated, their loss dependence on radius is analyzed, and fabricated bends have a loss in the range 0.8-1.6 dB per 90 degrees bend. Simulated and measured results show promise for large-scale fabrication of complex optical devices and high sensitivity sensors based on SWG waveguides with engineered optical properties, tailored to specific applications.

Silicon-on-insulator sensors using integrated resonance-enhanced defect-mediated photodetectors.

A resonance-enhanced, defect-mediated, ring resonator photodetector has been implemented as a single unit biosensor on a silicon-on-insulator platform, providing a cost effective means of integrating ring resonator sensors with photodetectors for lab-on-chip applications. This method overcomes the challenge of integrating hybrid photodetectors on the chip. The demonstrated responsivity of the photodetector-sensor was 90 mA/W. Devices were characterized using refractive index modified solutions and showed sensitivities of 30 nm/RIU.

Performance of ultra-thin SOI-based resonators for sensing applications.

This work presents simulation and experimental results of ultra-thin optical ring resonators, having larger Evanescent Field (EF) penetration depths, and therefore larger sensitivities, as compared to conventional Silicon-on-Insulator (SOI)-based resonator sensors. Having higher sensitivities to the changes in the refractive indices of the cladding media is desirable for sensing applications, as the interactions of interest take place in this region. Using ultra-thin waveguides (<100 nm thick) shows promise to enhance sensitivity for both bulk and surface sensing, due to increased penetration of the EF into the cladding. In this work, the designs and characterization of ultra-thin resonator sensors, within the constraints of a multi-project wafer service that offers three waveguide thicknesses (90 nm, 150 nm, and 220 nm), are presented. These services typically allow efficient integration of biosensors with on-chip detectors, moving towards the implementation of lab-on-chip (LoC) systems. Also, higher temperature stability of ultra-thin resonator sensors were characterized and, in the presence of intentional environmental (temperature) fluctuations, were compared to standard transverse electric SOI-based resonator sensors.

Silicon photonic micro-disk resonators for label-free biosensing.

Silicon photonic biosensors are highly attractive for multiplexed Lab-on-Chip systems. Here, we characterize the sensing performance of 3 µm TE-mode and 10 µm dual TE/TM-mode silicon photonic micro-disk resonators and demonstrate their ability to detect the specific capture of biomolecules. Our experimental results show sensitivities of 26 nm/RIU and 142 nm/RIU, and quality factors of 3.3x10(4) and 1.6x10(4) for the TE and TM modes, respectively. Additionally, we show that the large disks contain both TE and TM modes with differing sensing characteristics. Finally, by serializing multiple disks on a single waveguide bus in a CMOS compatible process, we demonstrate a biosensor capable of multiplexed interrogation of biological samples.

Oxygen Measurement in Microdevices.

Oxygen plays a fundamental role in respiration and metabolism, and quantifying oxygen levels is essential in many environmental, industrial, and research settings. Microdevices facilitate the study of dynamic, oxygen-dependent effects in real time. This review is organized around the key needs for oxygen measurement in microdevices, including integrability into microfabricated systems; sensor dynamic range and sensitivity; spatially resolved measurements to map oxygen over two- or three-dimensional regions of interest; and compatibility with multimodal and multianalyte measurements. After a brief overview of biological readouts of oxygen, followed by oxygen sensor types that have been implemented in microscale devices and sensing mechanisms, this review presents select recent applications in organs-on-chip in vitro models and new sensor capabilities enabling oxygen microscopy, bioprocess manufacturing, and pharmaceutical industries. With the advancement of multiplexed, interconnected sensors and instruments and integration with industry workflows, intelligent microdevice-sensor systems including oxygen sensors will have further impact in environmental science, manufacturing, and medicine.

Optical Attenuators Extend Dynamic Range but Alter Angular Response of Planar Ultraviolet-C Dosimeters.

Effective ultraviolet-C (UV-C) decontamination protocols of N95 respirators require validation that the entire N95 surface receives sufficient dose. Photochromic indicators (PCIs) can accurately measure UV-C dose on nonplanar surfaces, but often saturate below doses required to decontaminate porous, multilayered textiles like N95s. Here, we investigate the use of optical attenuators to extend PCI dynamic range while maintaining a near-ideal angular response-critical for accurate measurements of uncollimated UV-C. We show analytically that tuning attenuator refractive index, attenuation coefficient, and thickness can extend dynamic range, but compromises angular response unless the attenuator is an ideal diffuser. To investigate this tradeoff empirically, we stack PCIs behind model specular (floated borosilicate) and diffuse (polytetrafluoroethylene) attenuators, characterize the angular response, and evaluate on-N95 UV-C measurement accuracy within a decontamination system. Both attenuators increase PCI dynamic range >4×, but simultaneously introduce angle-dependent transmittance, which causes location-dependent underestimation of UV-C dose. PCI-borosilicate and PCI-polytetrafluoroethylene stacks underreport true on-N95 dose by (1) 14.7% and 3.6%, respectively, when near-normal to the source lamp array, and (2) 40.8% and 19.8%, respectively, in a steeply sloped location. Overall, we demonstrate that while planar attenuators can increase PCI dynamic range, verifying near-ideal angular response is critical for accurate UV-C measurements.

Biofunctionalization of Multiplexed Silicon Photonic Biosensors.

Silicon photonic (SiP) sensors offer a promising platform for robust and low-cost decentralized diagnostics due to their high scalability, low limit of detection, and ability to integrate multiple sensors for multiplexed analyte detection. Their CMOS-compatible fabrication enables chip-scale miniaturization, high scalability, and low-cost mass production. Sensitive, specific detection with silicon photonic sensors is afforded through biofunctionalization of the sensor surface; consequently, this functionalization chemistry is inextricably linked to sensor performance. In this review, we first highlight the biofunctionalization needs for SiP biosensors, including sensitivity, specificity, cost, shelf-stability, and replicability and establish a set of performance criteria. We then benchmark biofunctionalization strategies for SiP biosensors against these criteria, organizing the review around three key aspects: bioreceptor selection, immobilization strategies, and patterning techniques. First, we evaluate bioreceptors, including antibodies, aptamers, nucleic acid probes, molecularly imprinted polymers, peptides, glycans, and lectins. We then compare adsorption, bioaffinity, and covalent chemistries for immobilizing bioreceptors on SiP surfaces. Finally, we compare biopatterning techniques for spatially controlling and multiplexing the biofunctionalization of SiP sensors, including microcontact printing, pin- and pipette-based spotting, microfluidic patterning in channels, inkjet printing, and microfluidic probes.

PDMS Organ-On-Chip Design and Fabrication: Strategies for Improving Fluidic Integration and Chip Robustness of Rapidly Prototyped Microfluidic In Vitro Models.

The PDMS-based microfluidic organ-on-chip platform represents an exciting paradigm that has enjoyed a rapid rise in popularity and adoption. A particularly promising element of this platform is its amenability to rapid manufacturing strategies, which can enable quick adaptations through iterative prototyping. These strategies, however, come with challenges; fluid flow, for example, a core principle of organs-on-chip and the physiology they aim to model, necessitates robust, leak-free channels for potentially long (multi-week) culture durations. In this report, we describe microfluidic chip fabrication methods and strategies that are aimed at overcoming these difficulties; we employ a subset of these strategies to a blood-brain-barrier-on-chip, with others applied to a small-airway-on-chip. Design approaches are detailed with considerations presented for readers. Results pertaining to fabrication parameters we aimed to improve (e.g., the thickness uniformity of molded PDMS), as well as illustrative results pertaining to the establishment of cell cultures using these methods will also be presented.

An Optimization Framework for Silicon Photonic Evanescent-Field Biosensors Using Sub-Wavelength Gratings.

Silicon photonic (SiP) evanescent-field biosensors aim to combine the information-rich readouts offered by lab-scale diagnostics, at a significantly lower cost, and with the portability and rapid time to result offered by paper-based assays. While SiP biosensors fabricated with conventional strip waveguides can offer good sensitivity for label-free detection in some applications, there is still opportunity for improvement. Efforts have been made to design higher-sensitivity SiP sensors with alternative waveguide geometries, including sub-wavelength gratings (SWGs). However, SWG-based devices are fragile and prone to damage, limiting their suitability for scalable and portable sensing. Here, we investigate SiP microring resonator sensors designed with SWG waveguides that contain a "fishbone" and highlight the improved robustness offered by this design. We present a framework for optimizing fishbone-style SWG waveguide geometries based on numerical simulations, then experimentally measure the performance of ring resonator sensors fabricated with the optimized waveguides, targeting operation in the O-band and C-band. For the O-band and C-band devices, we report bulk sensitivities up to 349 nm/RIU and 438 nm/RIU, respectively, and intrinsic limits of detection as low as 5.1 × 10-4 RIU and 7.1 × 10-4 RIU, respectively. This performance is comparable to the state of the art in SWG-based sensors, positioning fishbone SWG resonators as an attractive, more robust, alternative to conventional SWG designs.

Current Understanding of Ultraviolet-C Decontamination of N95 Filtering Facepiece Respirators.

Introduction: The COVID-19 pandemic has led to critical shortages of single-use N95 filtering facepiece respirators. The US Centers for Disease Control and Prevention has identified ultraviolet-C (UV-C) irradiation as one of the most promising decontamination methods during crisis-capacity surges; however, understanding the mechanism of pathogen inactivation and post-treatment respirator performance is central to effective UV-C decontamination. Objective: We summarize the UV-C N95 decontamination evidence and identify key metrics. Methods: We evaluate the peer-reviewed literature on UV-C decontamination to inactivate SARS-CoV-2, viral analogues, and other microorganisms inoculated on N95s, as well as the resulting effect on respirator fit and filtration. Where peer-reviewed studies are absent, we discuss outstanding questions and ongoing work. Key Findings: Evidence supports that UV-C exposure of ≥1.0 J/cm2 inactivates SARS-CoV-2 analogues (≥3-log reduction) on the majority of tested N95 models. The literature cautions that (1) viral inactivation is N95 model-dependent and impeded by shadowing, (2) N95 straps require secondary decontamination, (3) higher doses may be necessary to inactivate other pathogens (e.g., some bacterial spores), and (4) while N95 fit and filtration appear to be preserved for 10-20 cycles of 1.0 J/cm2, donning and doffing may degrade fit to unacceptable levels within fewer cycles. Results and Discussion: Effective N95 UV-C treatment for emergency reuse requires both (1) inactivation of the SARS-CoV-2 virus, achieved through application of UV-C irradiation at an appropriate wavelength and effective dose, and (2) maintenance of the fit and filtration efficiency of the N95. Conclusions: UV-C treatment is a risk-mitigation process that should be implemented only under crisis-capacity conditions and with proper engineering, industrial hygiene, and biosafety controls.

Quantitative UV-C dose validation with photochromic indicators for informed N95 emergency decontamination.

With COVID-19 N95 shortages, frontline medical personnel are forced to reuse this disposable-but sophisticated-multilayer respirator. Widely used to decontaminate nonporous surfaces, UV-C light has demonstrated germicidal efficacy on porous, non-planar N95 respirators when all surfaces receive ≥1.0 J/cm2 dose. Of utmost importance across disciplines, translation of empirical evidence to implementation relies upon UV-C measurements frequently confounded by radiometer complexities. To enable rigorous on-respirator measurements, we introduce a photochromic indicator dose quantification technique for: (1) UV-C treatment design and (2) in-process UV-C dose validation. While addressing outstanding indicator limitations of qualitative readout and insufficient dynamic range, our methodology establishes that color-changing dosimetry can achieve the necessary accuracy (>90%), uncertainty (<10%), and UV-C specificity (>95%) required for UV-C dose measurements. In a measurement infeasible with radiometers, we observe a striking ~20× dose variation over N95s within one decontamination system. Furthermore, we adapt consumer electronics for accessible quantitative readout and use optical attenuators to extend indicator dynamic range >10× to quantify doses relevant for N95 decontamination. By transforming photochromic indicators into quantitative dosimeters, we illuminate critical considerations for both photochromic indicators themselves and UV-C decontamination processes.

Mapping of UV-C dose and SARS-CoV-2 viral inactivation across N95 respirators during decontamination.

During public health crises like the COVID-19 pandemic, ultraviolet-C (UV-C) decontamination of N95 respirators for emergency reuse has been implemented to mitigate shortages. Pathogen photoinactivation efficacy depends critically on UV-C dose, which is distance- and angle-dependent and thus varies substantially across N95 surfaces within a decontamination system. Due to nonuniform and system-dependent UV-C dose distributions, characterizing UV-C dose and resulting pathogen inactivation with sufficient spatial resolution on-N95 is key to designing and validating UV-C decontamination protocols. However, robust quantification of UV-C dose across N95 facepieces presents challenges, as few UV-C measurement tools have sufficient (1) small, flexible form factor, and (2) angular response. To address this gap, we combine optical modeling and quantitative photochromic indicator (PCI) dosimetry with viral inactivation assays to generate high-resolution maps of "on-N95" UV-C dose and concomitant SARS-CoV-2 viral inactivation across N95 facepieces within a commercial decontamination chamber. Using modeling to rapidly identify on-N95 locations of interest, in-situ measurements report a 17.4 ± 5.0-fold dose difference across N95 facepieces in the chamber, yielding 2.9 ± 0.2-log variation in SARS-CoV-2 inactivation. UV-C dose at several on-N95 locations was lower than the lowest-dose locations on the chamber floor, highlighting the importance of on-N95 dose validation. Overall, we integrate optical simulation with in-situ PCI dosimetry to relate UV-C dose and viral inactivation at specific on-N95 locations, establishing a versatile approach to characterize UV-C photoinactivation of pathogens contaminating complex substrates such as N95s.

Optical oxygen sensors for applications in microfluidic cell culture.

The presence and concentration of oxygen in biological systems has a large impact on the behavior and viability of many types of cells, including the differentiation of stem cells or the growth of tumor cells. As a result, the integration of oxygen sensors within cell culture environments presents a powerful tool for quantifying the effects of oxygen concentrations on cell behavior, cell viability, and drug effectiveness. Because microfluidic cell culture environments are a promising alternative to traditional cell culture platforms, there is recent interest in integrating oxygen-sensing mechanisms with microfluidics for cell culture applications. Optical, luminescence-based oxygen sensors, in particular, show great promise in their ability to be integrated with microfluidics and cell culture systems. These sensors can be highly sensitive and do not consume oxygen or generate toxic byproducts in their sensing process. This paper presents a review of previously proposed optical oxygen sensor types, materials and formats most applicable to microfluidic cell culture, and analyzes their suitability for this and other in vitro applications.

Rapid electrotransfer probing for improved detection sensitivity in in-gel immunoassays.

Protein electrotransfer in conventional western blotting facilitates detection of size-separated proteins by diffusive immunoprobing, as analytes are transferred from a small-pore sizing gel to a blotting membrane for detection. This additional transfer step can, however, impair detection sensitivity through protein losses and confound protein localization. To overcome challenges associated with protein transfer, in-gel immunoassays immobilize target proteins to the hydrogel matrix for subsequent in-gel immunoprobing. Yet, detection sensitivity in diffusive immunoprobing of hydrogels is determined by the gel pore size relative to the probe size, and in-gel immunoprobing results in (i) reduced in-gel probe concentration compared to surrounding free-solution, and (ii) slow in-gel probe transfer compared to immunocomplex dissociation. Here, we demonstrate electrotransfer probing for effective and rapid immunoprobing of in-gel immunoassays. Critically, probe (rather than target protein) is electrotransferred from an inert, large-pore 'loading gel' to a small-pore protein sizing gel. Electric field is used as a tuneable parameter for electromigration velocity, providing electrotransfer probing with a fundamental advantage over diffusive probing. Using electrotransfer probing, we observe 6.5 ± 0.1× greater probe concentration loaded in-gel in ∼82× time reduction, and 2.7 ± 0.4× less probe concentration remaining in-gel after unloading in ∼180× time reduction (compared to diffusive probing). We then apply electrotransfer probing to detect OVA immobilized in-gel and achieve 4.1 ± 3.4× greater signal-to-noise ratio and 30× reduction in total immunoprobing duration compared to diffusive probing. We demonstrate electrotransfer probing as a substantially faster immunoprobing method for improved detection sensitivity of protein sizing in-gel immunoassays.

3D projection electrophoresis for single-cell immunoblotting.

Immunoassays and mass spectrometry are powerful single-cell protein analysis tools; however, interfacing and throughput bottlenecks remain. Here, we introduce three-dimensional single-cell immunoblots to detect both cytosolic and nuclear proteins. The 3D microfluidic device is a photoactive polyacrylamide gel with a microwell array-patterned face (xy) for cell isolation and lysis. Single-cell lysate in each microwell is "electrophoretically projected" into the 3rd dimension (z-axis), separated by size, and photo-captured in the gel for immunoprobing and confocal/light-sheet imaging. Design and analysis are informed by the physics of 3D diffusion. Electrophoresis throughput is > 2.5 cells/s (70× faster than published serial sampling), with 25 immunoblots/mm2 device area (>10× increase over previous immunoblots). The 3D microdevice design synchronizes analyses of hundreds of cells, compared to status quo serial analyses that impart hours-long delay between the first and last cells. Here, we introduce projection electrophoresis to augment the heavily genomic and transcriptomic single-cell atlases with protein-level profiling.