multimodal-analysis-microplastics-drinking-water-using-silicon

Multimodal Analysis of Microplastics in Drinking Water using a Silicon Nanomembrane Analysis Pipeline

The biological impact of microplastic pollution in human food and water sources is largely unknown, and drinking water sources are not exempt from this ...

SiMPore Inc.

Imaging with Zernike phase plates is increasingly being used in cryo-TEM tomography and cryo-EM single-particle applications. However, rapid ageing of the phase plates, together with the cost and effort in producing them, present serious obstacles to widespread adoption. We are experimenting with phase plates based on silicon chips that have thin windows; such phase plates could be mass-produced and made available at moderate cost. The windows are coated with conductive layers to reduce charging, and this considerably extends the useful life of the phase plates compared to traditional pure-carbon phase plates. However, a compromise must be reached between robustness and transmission through the phase-plate film. Details are given on testing phase-plate performance by means of imaging an amorphous thin film and evaluating the power spectra of the images.

Methods for testing Zernike phase plates and a report on silicon-based phase plates with reduced charging and improved ageing characteristics.

Typical in vitro barrier and co-culture models rely upon thick semi-permeable polymeric membranes that physically separate two compartments. Polymeric track-etched membranes, while permeable to small molecules, are far from physiological with respect to physical interactions with co-cultured cells and are not compatible with high-resolution imaging due to light scattering and autofluorescence. Here we report on an optically transparent ultrathin membrane with porosity exceeding 20%. We optimize deposition and annealing conditions to create a tensile and robust porous silicon dioxide membrane that is comparable in thickness to the vascular basement membrane (100-300 nm). We demonstrate that human umbilical vein endothelial cells (HUVECs) spread and proliferate on these membranes similarly to control substrates. Additionally, HUVECs are able to transfer cytoplasmic cargo to adipose-derived stem cells when they are co-cultured on opposite sides of the membrane, demonstrating its thickness supports physiologically relevant cellular interactions. Lastly, we confirm that these porous glass membranes are compatible with lift-off processes yielding membrane sheets with an active area of many square centimeters. We believe that these membranes will enable new in vitro barrier and co-culture models while offering dramatically improved visualization compared to conventional alternatives.

Ultrathin transparent membranes for cellular barrier and co-culture models.

The widely used 0.2/0.22 µm polymer sterile filters were developed for small molecule and protein sterile filtration but are not well-suited for the production of large nonprotein biological therapeutics, resulting in significant yield loss and production cost increases. Here, we report on the development of membranes with isoporous sub-0.2 μm rectangular prism pores using silicon micromachining to produce microslit silicon nitride (MSN) membranes. The very high porosity (~33%) and ultrathin (200 nm) nature of the 0.2 µm MSN membranes results in a dramatically different structure than the traditional 0.2/0.22 µm polymer sterile filter, which yielded comparable performance properties (including gas and hydraulic permeance, maximum differential pressure tolerance, nanoparticle sieving/fouling behavior). The results from bacteria retention tests, conducted according to the guidance of regulatory agencies, demonstrated that the 0.2 µm MSN membranes can be effectively used as sterile filters. It is anticipated that the results and technologies presented in this study will find future utility in the production of non-protein biological therapeutics and in other biological and biomedical applications.

Development of isoporous microslit silicon nitride membranes for sterile filtration applications.

Nanoscale preconfinement of DNA has been shown to reduce the variation of passage times through solid-state nanopores. Preconfinement has been previously achieved by forming a femtoliter-sized cavity capped with a highly porous layer of nanoporous silicon nitride (NPN). This cavity was formed by sealing a NPN nanofilter membrane against a substrate chip using water vapor delamination. Ultimately, this method of fabrication cannot keep a consistent spacing between the filter and solid-state nanopore due to thermal fluctuations and wrinkles in the membrane, nor can it be fabricated on thousands of individual devices reliably. To overcome these issues, we present a method to fabricate the femtoliter cavity monolithically, using a selective XeF etch to hollow out a polysilicon spacer sandwiched between silicon nitride layers. These monolithically fabricated cavities behave identically to their counterparts formed by vapor delamination, exhibiting similar translocation passage time variation reduction and folding suppression of DNA without requiring extensive manual assembly. The ability to form nanocavity sensors with nanometer-scale precision and to reliably manufacture them at scale using batch wafer processing techniques will find numerous applications, including motion control of polymers for single-molecule detection applications, filtering of dirty samples prior to nanopore detection, and simple fabrication of single-molecule nanobioreactors.

Monolithic Fabrication of NPN/SiN Dual Membrane Cavity for Nanopore-based DNA Sensing.

Pollution by microplastics (MPs) is a growing problem that is now well-recognized, as concerning levels of MPs have been found in drinking water, food, and even human tissues. Given the evolving understanding of their toxicological effects on human health, MPs are an area of concern requiring further study. Consequently, there is a need for greater understanding of the performance characteristics of common MP analytical methods and where possible, for standardizing methods and reporting practices. Here, we report our work comparing filtration and imaging properties of five analytical filters suitable for MP capture and analysis. We compared track-etched polycarbonate with (PCTEG) and without gold coating (PCTE), polytetrafluoroethylene (PTFE), porous silicon (PSi), and gold-coated microslit silicon nitride membranes (MSSN-Au). Four of the filter types had a nominal 1.0 μm cut-off, except for PCTEG which had a 0.8 nominal cut-off. We examined the ultrastructure of each membrane type by electron microscopy to understand how their physical properties influence filtration and imaging performance. We compared clean water filtration rates and timed volume passage for each filter in comparison to its porosity and working surface area. We further compared optical microscopy imaging properties for each filter with model MP samples in both bright-field and fluorescent modes with accompanying Nile Red staining. In terms of absolute and surface area-normalized flow rates, our measurements ranked the filters in order of MSSN-Au > PTFE > PCTE > PCTEG > PSi. Similarly, we found MSSN-Au filters compared favorably in terms of optical microscopy performance. Collectively, these data will aid practitioners when choosing analytical filters for MP surveillance and testing.

Article	Year
Methods for testing Zernike phase plates and a report on silicon-based phase plates with reduced charging and improved ageing characteristics. Journal of structural biology\| PubMed ID: 23994351	2013
Ultrathin transparent membranes for cellular barrier and co-culture models. Biofabrication\| PubMed ID: 28140345	2017
Development of isoporous microslit silicon nitride membranes for sterile filtration applications. Biotechnology and bioengineering\| PubMed ID: 31784974	2020
Monolithic Fabrication of NPN/SiN Dual Membrane Cavity for Nanopore-based DNA Sensing. Advanced materials interfaces\| PubMed ID: 32577337	2019
Comparative evaluation of filtration and imaging properties of analytical filters for microplastic capture and analysis. Chemosphere\| PubMed ID: 37127196	2023

James Roussie

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