25 research outputs found
Justify your alpha
Benjamin et al. proposed changing the conventional “statistical significance” threshold (i.e.,the alpha level) from p ≤ .05 to p ≤ .005 for all novel claims with relatively low prior odds. They provided two arguments for why lowering the significance threshold would “immediately improve the reproducibility of scientific research.” First, a p-value near .05provides weak evidence for the alternative hypothesis. Second, under certain assumptions, an alpha of .05 leads to high false positive report probabilities (FPRP2 ; the probability that a significant finding is a false positive
Justify your alpha
In response to recommendations to redefine statistical significance to p ≤ .005, we propose that researchers should transparently report and justify all choices they make when designing a study, including the alpha level
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.Applied Science, Faculty ofElectrical and Computer Engineering, Department ofReviewedFacult
Recommended from our members
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
Recommended from our members
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
Microfluidic cell culture systems with integrated sensors for drug screening.
Cell-based testing is a key step in drug screening for cancer treatments. A microfluidic platform
can permit more precise control of the cell culture microenvironment, such as gradients in soluble
factors. These small-scale devices also permit tracking of low cell numbers. As a new screening
paradigm, a microscale system for integrated cell culture and drug screening promises to provide a
simple, scalable tool to apply standardized protocols used in cellular response assays. With the
ability to dynamically control the microenvironment, we can create temporally varying drug profiles
to mimic physiologically measured profiles. In addition, low levels of oxygen in cancerous tumors
have been linked with drug resistance and decreased likelihood of successful treatment and patient
survival. Our work also integrates a thin-film oxygen sensor with a microfluidic oxygen gradient
generator which will in future allow us to create spatial oxygen gradients and study effects of
hypoxia on cell response to drug treatment. In future, this technology promises to improve cell-based
validation in the drug discovery process, decreasing the cost and increasing the speed in screening
large numbers of compounds.
Copyright 2012 Society of Photo-Optical Instrumentation Engineers.
One print or electronic copy may be made for personal use only. Systematic reproduction and distribution,
duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper are prohibited.Applied Science, Faculty ofElectrical and Computer Engineering, Department ofReviewedFacult
Cascaded silicon-on-insulator microring resonators for the detection of biomolecules in PDMS microfluidic channel
Silicon-On-Insulator (SOI) photonic microring resonators have shown promising potential for real time detection of biomolecules because of the
sensitivity towards surface binding events. Previous work shows the use of single ring resonators for sensing applications. Each ring requires
an input and output coupler and can be addressed only one at a time. We propose a novel use of cascaded ring resonators (width w = 200 nm and
bending Radius R = 30 µm) together with a PDMS microfluidic network fabricated by soft lithography to expose each ring individually with
different solutions. The SOI substrate with the planar waveguides and the PDMS with the microchannels are reversibly bonded to each other. The
use of cascaded ring resonators offers the possibility to measure transmission spectra of multiple rings in different channels simultaneously.
We measured Q-factors of >30'000 in air and >10'000 when exposed to water. Using a water/glycerin solution with known refractive indices we
determine the sensitivity to be ~40 nm/RIU.
Copyright 2011 Society of Photo-Optical Instrumentation Engineers.
One print or electronic copy may be made for personal use only. Systematic reproduction and distribution,
duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper are prohibited.Applied Science, Faculty ofElectrical and Computer Engineering, Department ofReviewedFacult
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 (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
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.Applied Science, Faculty ofElectrical and Computer Engineering, Department ofReviewedFacult