Ole Behrmann

MEMS environmental sensors, including pressure, gas, and humidity sensors, require protection from mechanical damage, particle exposure, and condensing moisture, while maintaining their ability to exchange gases with the environment. This work introduces a novel packaging approach for MEMS environmental sensors using substrate-embedded filters made from microfine powders through PowderMEMS^® microfabrication technology. The study demonstrates the successful fabrication of gas permeable, functionalized PowderMEMS^® filters on 200 mm Si-wafers for wafer-level packaging of MEMS environmental sensors. Utilizing complete Si-wafers allows for all MEMS sensors on a device wafer to be packaged in a single substrate bonding step, followed by die singulation. The processed wafers are shown to be compatible with high-temperature glass-frit substrate bonding. Alternatively, individual chips with PowderMEMS^® filters can be assembled discretely onto standard semiconductor packages to serve as gas-permeable filters. Successful hydrophobation of the inherently hydrophilic PowderMEMS^® structures by deposition of hydrophobic nanofilms is demonstrated and resistance to water ingress is evaluated by immersion testing. Given that many MEMS gas sensors are cross-reactive to oxidizing gases like ozone, this study also explores the integration of ozone-degrading catalytic powder into the PowderMEMS^® filters. As a proof-of-concept, commercial MEMS ozone sensors are modified with catalytic PowderMEMS^® caps, and successful ozone degradation is demonstrated. While PowderMEMS^® processing is typically conducted on 200 mm Si-wafers, other suitable substrates include glass and (fiber-reinforced) polymers.

This work presents new porous environmental protection caps designed for wafer level chip scale packaging (WLCSP) of MEMS environmental sensors. The caps consist of gaspermeable microstructures formed from loose aluminum oxide powder solidified by a ceramic thin film, grown using atomic layer deposition (ALD). For the first time, the full process flow of the proposed approach is demonstrated by manufacture of the cap wafer followed by substrate bonding using glass-frit technology. By analyzing the influence of the caps on the response time of MEMS humidity sensors, this study proves the viability of the proposed packaging technology. In addition, the ability to further functionalize the porous caps is demonstrated by the deposition of a superhydrophobic polymer thin film by chemical vapor deposition.

Most current thermal MEMS use fragile structures such as thin-film membranes or microcantilevers for thermal isolation. To increase the robustness of these devices, solid thermal insulators that are compatible with MEMS cleanroom processing are needed. This work introduces a novel approach for microscale thermal isolation using porous microstructures created with the recently developed PowderMEMS wafer-level process. MEMS devices consisting of heaters on a thin-film membrane were modified with porous microstructures made from three different materials. A thermal model for the estimation of the resulting thermal conductivity was developed, and measurements for porous structures in ambient air and under vacuum were performed. The PowderMEMS process was successfully used to create microscale thermal insulators in silicon cavities at the wafer level. Measurements indicate thermal conductivities of close to 0.1 W/mK in ambient air and close to 0.04 W/mK for porous structures under vacuum for the best-performing material. The obtained thermal conductivities are lower than those reported for both glass and porous silicon, making PowderMEMS a very interesting alternative for solid microscale thermal isolation.

A comprehensive overview of PowderMEMS—a novel back-end-of-line-compatible microfabrication technology—is presented in this paper. The PowderMEMS process solidifies micron-sized particles via atomic layer deposition (ALD) to create three-dimensional microstructures on planar substrates from a wide variety of materials. The process offers numerous degrees of freedom for the design of functional MEMSs, such as a wide choice of different material properties and the precise definition of 3D volumes at the substrate level, with a defined degree of porosity. This work details the characteristics of PowderMEMS materials as well as the maturity of the fabrication technology, while highlighting prospects for future microdevices. Applications of PowderMEMS in the fields of magnetic, thermal, optical, fluidic, and electrochemical MEMSs are described, and future developments and challenges of the technology are discussed.

In March 2020, the SARS-CoV-2 virus outbreak was declared as a world pandemic by the World Health Organization (WHO). The only measures for controlling the outbreak are testing and isolation of infected cases. Molecular real-time polymerase chain reaction (PCR) assays are very sensitive but require highly equipped laboratories and well-trained personnel. In this study, a rapid point-of-need detection method was developed to detect the RNA-dependent RNA polymerase (RdRP), envelope protein (E), and nucleocapsid protein (N) genes of SARS-CoV-2 based on the reverse transcription recombinase polymerase amplification (RT-RPA) assay. RdRP, E, and N RT-RPA assays required approximately 15 min to amplify 2, 15, and 15 RNA molecules of molecular standard/reaction, respectively. RdRP and E RT-RPA assays detected SARS-CoV-1 and 2 genomic RNA, whereas the N RT-RPA assay identified only SARS-CoV-2 RNA. All established assays did not cross-react with nucleic acids of other respiratory pathogens. The RT-RPA assay’s clinical sensitivity and specificity in comparison to real-time RT-PCR (n = 36) were 94 and 100% for RdRP; 65 and 77% for E; and 83 and 94% for the N RT-RPA assay. The assays were deployed to the field, where the RdRP RT-RPA assays confirmed to produce the most accurate results in three different laboratories in Africa (n = 89). The RPA assays were run in a mobile suitcase laboratory to facilitate the deployment at point of need. The assays can contribute to speed up the control measures as well as assist in the detection of COVID-19 cases in low-resource settings.

We investigate the compatibility of three 3D printing materials towards real-time recombinase polymerase amplification (rtRPA). Both the general ability of the rtRPA reaction to occur while in contact with the cured 3D printing materials as well as the residual autofluorescence and fluorescence drift in dependence on post curing of the materials is characterized. We 3D printed monolithic rtRPA microreactors and subjected the devices to different post curing protocols. Residual autofluorescence and drift, as well as rtRPA kinetics, were then measured in a custom-made mobile temperature-controlled fluorescence reader (mTFR). Furthermore, we investigated the effects of storage on the devices over a 30-day period. Finally, we present the single- and duplex rtRPA detection of both the organism-specific Klebsiella haemolysin (khe) gene and the New Delhi metallo-β-lactamase 1 (bla_NDM-1) gene from Klebsiella pneumoniae. Results: No combination of 3D printing resin and post curing protocol completely inhibited the rtRPA reaction. The autofluorescence and fluorescence drift measured were found to be highly dependent on printing material and wavelength. Storage had the effect of decreasing the autofluorescence of the investigated materials. Both khe and bla_NDM-1 were successfully detected by single- and duplex-rtRPA inside monolithic rtRPA microreactors printed from NextDent Ortho Clear (NXOC). The reaction kinetics were found to be close to those observed for rtRPA performed in a microcentrifuge tube without the need for mixing during amplification. Singleplex assays for both khe and bla_NDM-1 achieved a limit of detection of 2.5 × 10¹ DNA copies while the duplex assay achieved 2.5 × 10¹ DNA copies for khe and 2.5 × 10² DNA copies for bla_NDM-1. Impact: We expand on the state of the art by demonstrating a technology that can manufacture monolithic microfluidic devices that are readily suitable for rtRPA. The devices exhibit very low autofluorescence and fluorescence drift and are compatible with RPA chemistry without the need for any surface pre-treatment such as blocking with, e.g., BSA or PEG.

Background: The current outbreak of SARS-CoV-2 has spread to almost every country with more than 5 million confirmed cases and over 300,000 deaths as of May 26, 2020. Rapid first-line testing protocols are needed for outbreak control and surveillance. Methods: We used computational and manual designs to generate a suitable set of reverse transcription recombinase polymerase amplification (RT-RPA) primer and exonuclease probe, internally quenched (exo-IQ), sequences targeting the SARS-CoV-2 N gene. RT-RPA sensitivity was determined by amplification of in vitro transcribed RNA standards. Assay selectivity was demonstrated with a selectivity panel of 32 nucleic acid samples derived from common respiratory viruses. To validate the assay against full-length SARS-CoV-2 RNA, total viral RNA derived from cell culture supernatant and 19 nasopharyngeal swab samples (8 positive and 11 negative for SARS-CoV-2) were screened. All results were compared to established RT-qPCR assays. Results: The 95% detection probability of the RT-RPA assay was determined to be 7.74 (95% CI: 2.87–27.39) RNA copies per reaction. The assay showed no cross-reactivity to any other screened coronaviruses or respiratory viruses of clinical significance. The developed RT-RPA assay produced 100% diagnostic sensitivity and specificity when compared to RT-qPCR (n = 20). Conclusions: With a run time of 15 to 20 minutes and first results being available in under 7 minutes for high RNA concentrations, the reported assay constitutes one of the fastest nucleic acid based detection methods for SARS-CoV-2 to date and may provide a simple-to-use alternative to RT-qPCR for first-line screening at the point of need.

Nucleic acid amplification techniques such as real-time PCR are essential instruments for the identification and quantification of viruses. They are fast, very sensitive and highly specific, but often require elaborate and labor intensive sample preparation to achieve successful amplification of the target sequence. In this work we demonstrate the complete microfluidic preparation of amplifiable virus DNA from dilute specimens. Our approach combines free-flow electrophoretic preconcentration of viral particles with thermal lysis and gel-electrophoretic nucleic acid extraction on a single device. The on-chip preconcentration achieves a capture efficiency of >99% for dilute suspensions of bacteriophage PhiX174. Following preconcentration, phages are thermally lysed and released DNA is recovered after 40 s of on-chip gel-electrophoresis with a recovery rate of ∼73%. Furthermore we demonstrate a detection limit of ∼1 PFU ml⁻¹ (∼0.02 DNA copies per μl) for the detection of bacteriophage PhiX174 by PCR. To simplify operation of the device, we describe the development of a custom-made chip holder as well as a compact peristaltic pump and power supply, which enable user-friendly operation with low risk of cross-contamination and high potential for automation in the field of point-of-care diagnostics.