scholarly journals CellProfiler: A fit tool for image analysis in droplet microfluidics

2019 ◽  
Author(s):  
Simona Bartkova ◽  
Marko Vendelin ◽  
Immanuel Sanka ◽  
Pille Pata ◽  
Ott Scheler

AbstractDroplet microfluidic assays are rapidly gaining popularity as the result of the ability to manipulate and monitor single biological molecules, individual cells or small populations of bacteria in pico- and nanoliter droplets, with high sensitivity, precision and accuracy in a high-throughput manner. Nonetheless, there is a demand for user-friendly and low-cost droplet analysis technology. In this article we meet this demand with free open-source software CellProfiler (CP). To illustrate the competence of CP as a droplet analysis tool, we show droplet digital quantification of viable fluorescent bacteria.

Research ◽  
2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Yang Wang ◽  
Kaiju Li ◽  
Gaolian Xu ◽  
Chuan Chen ◽  
Guiqin Song ◽  
...  

Sensitive detection of SARS-CoV-2 is of great importance for inhibiting the current pandemic of COVID-19. Here, we report a simple yet efficient platform integrating a portable and low-cost custom-made detector and a novel microwell array biochip for rapid and accurate detection of SARS-CoV-2. The instrument exhibits expedited amplification speed that enables colorimetric read-out within 25 minutes. A polymeric chip with a laser-engraved microwell array was developed to process the reaction between the primers and the respiratory swab RNA extracts, based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP). To achieve clinically acceptable performance, we synthesized a group of six primers to identify the conserved regions of the ORF1ab gene of SARS-CoV-2. Clinical trials were conducted with 87 PCR-positive and 43 PCR-negative patient samples. The platform demonstrated both high sensitivity (95.40%) and high specificity (95.35%), showing potentials for rapid and user-friendly diagnosis of COVID-19 among many other infectious pathogens.


2017 ◽  
Author(s):  
Peng Hu ◽  
Emily Fabyanic ◽  
Zhaolan Zhou ◽  
Hao Wu

Massively parallel single-cell RNA sequencing can precisely resolve cellular diversity in a high-throughput manner at low cost, but unbiased isolation of intact single cells from complex tissues, such as adult mammalian brains, is challenging. Here, we integrate sucrose-gradient assisted nuclear purification with droplet microfluidics to develop a highly scalable single-nucleus RNA-Seq approach (sNucDrop-Seq), which is free of enzymatic dissociation and nucleus sorting. By profiling ~11,000 nuclei isolated from adult mouse cerebral cortex, we demonstrate that sNucDrop-Seq not only accurately reveals neuronal and non-neuronal subtype composition with high sensitivity, but also enables analysis of long non-coding RNAs and transient states such as neuronal activity-dependent transcription at single-cell resolution in vivo.


Micromachines ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 481 ◽  
Author(s):  
Rumaner ◽  
Horowitz ◽  
Ovadya ◽  
Folch

In this paper we describe the use of thread as a low-cost material for a microfluidic chemosensitivity assay that uses intact tumor tissue ex vivo. Today, the need for new and effective cancer treatments is greater than ever, but unfortunately, the cost of developing new chemotherapy drugs has never been higher. Implementation of low-cost microfluidic techniques into drug screening devices could potentially mitigate some of the immense cost of drug development. Thread is an ideal material for use in drug screening as it is inexpensive, widely available, and can transport liquid without external pumping hardware, i.e., via capillary action. We have developed an inexpensive microfluidic delivery prototype that uses silk threads to selectively deliver fluids onto subregions of living xenograft tumor slices. Our device can be fabricated completely for less than $0.25 in materials and requires no external equipment to operate. We found that by varying thread materials, we could optimize device characteristics, such as flow rate; we specifically explored the behavior of silk, nylon, cotton, and polyester. The incremental cost of our device is insignificant compared to the tissue culture supplies. The use of thread as a microfluidic material has the potential to produce inexpensive, accessible, and user-friendly devices for drug testing that are especially suited for low-resource settings.


2021 ◽  
Author(s):  
Immanuel Sanka ◽  
Simona Bartkova ◽  
Pille Pata ◽  
Olli-Pekka Smolander ◽  
Ott Scheler

ABSTRACTDroplet microfluidics has revealed innovative strategies in biology and chemistry. This advancement has delivered novel quantification methods, such as digital droplet polymerase chain reaction (ddPCR) and antibiotic heteroresistance analysis tool. For droplet analysis, researchers often use imaging techniques. Unfortunately, the analysis of images may require specific tools or programming skills to produce the expected results. In order to address the issue, we explore the potential use of standalone freely available software to detect droplets. We select four most popular software and classify them into instinctive and objective types based on their operation logic. We test and evaluate the software’s i) ability to detect droplets, ii) accuracy and precision, and iii) overall user-friendliness. In our experimental setting we find the objective type of software is better suited for droplet detection. The objective type of software also has simpler workflow or pipeline, especially aimed for non-experienced user. In our case, CellProfiler™ (CP) offers the most user-friendly experience for both single image and batch processing analysis.


Sensors ◽  
2021 ◽  
Vol 21 (5) ◽  
pp. 1673
Author(s):  
Catherine Grogan ◽  
Faolan Radford McGovern ◽  
Rory Staines ◽  
George Amarandei ◽  
Izabela Naydenova

High-sensitivity and simple, low-cost readout are desirable features for sensors independent of the application area. Micro-cantilever sensors use the deflection induced by the analyte presence to achieve high-sensitivity but possess complex electronic readouts. Current holographic sensors probe the analyte presence by measuring changes in their optical properties, have a simpler low-cost readout, but their sensitivity can be further improved. Here, the two working principles were combined to obtain a new hybrid sensor with enhanced sensitivity. The diffractive element, a holographically patterned thin photopolymer layer, was placed on a polymer (polydimethylsiloxane) layer forming a bi-layer macro-cantilever. The different responses of the layers to analyte presence lead to cantilever deflection. The sensitivity and detection limits were evaluated by measuring the variation in cantilever deflection and diffraction efficiency with relative humidity. It was observed that the sensitivity is tunable by controlling the spatial frequency of the photopolymer gratings and the cantilever thickness. The sensor deflection was also visible to the naked eye, making it a simple, user-friendly device. The hybrid sensor diffraction efficiency response to the target analyte had an increased sensitivity (10-fold when compared with the cantilever or holographic modes operating independently), requiring a minimum upturn in the readout complexity.


2012 ◽  
Vol 44 (2) ◽  
pp. 75-93
Author(s):  
Peter Mortensen

This essay takes its cue from second-wave ecocriticism and from recent scholarly interest in the “appropriate technology” movement that evolved during the 1960s and 1970s in California and elsewhere. “Appropriate technology” (or AT) refers to a loosely-knit group of writers, engineers and designers active in the years around 1970, and more generally to the counterculture’s promotion, development and application of technologies that were small-scale, low-cost, user-friendly, human-empowering and environmentally sound. Focusing on two roughly contemporary but now largely forgotten American texts Sidney Goldfarb’s lyric poem “Solar-Heated-Rhombic-Dodecahedron” (1969) and Gurney Norman’s novel Divine Right’s Trip (1971)—I consider how “hip” literary writers contributed to eco-technological discourse and argue for the 1960s counterculture’s relevance to present-day ecological concerns. Goldfarb’s and Norman’s texts interest me because they conceptualize iconic 1960s technologies—especially the Buckminster Fuller-inspired geodesic dome and the Volkswagen van—not as inherently alienating machines but as tools of profound individual, social and environmental transformation. Synthesizing antimodernist back-to-nature desires with modernist enthusiasm for (certain kinds of) machinery, these texts adumbrate a humanity- and modernity-centered post-wilderness model of environmentalism that resonates with the dilemmas that we face in our increasingly resource-impoverished, rapidly warming and densely populated world.


2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Yehe Liu ◽  
Andrew M. Rollins ◽  
Richard M. Levenson ◽  
Farzad Fereidouni ◽  
Michael W. Jenkins

AbstractSmartphone microscopes can be useful tools for a broad range of imaging applications. This manuscript demonstrates the first practical implementation of Microscopy with Ultraviolet Surface Excitation (MUSE) in a compact smartphone microscope called Pocket MUSE, resulting in a remarkably effective design. Fabricated with parts from consumer electronics that are readily available at low cost, the small optical module attaches directly over the rear lens in a smartphone. It enables high-quality multichannel fluorescence microscopy with submicron resolution over a 10× equivalent field of view. In addition to the novel optical configuration, Pocket MUSE is compatible with a series of simple, portable, and user-friendly sample preparation strategies that can be directly implemented for various microscopy applications for point-of-care diagnostics, at-home health monitoring, plant biology, STEM education, environmental studies, etc.


Author(s):  
Antonia Perju ◽  
Nongnoot Wongkaew

AbstractLateral flow assays (LFAs) are the best-performing and best-known point-of-care tests worldwide. Over the last decade, they have experienced an increasing interest by researchers towards improving their analytical performance while maintaining their robust assay platform. Commercially, visual and optical detection strategies dominate, but it is especially the research on integrating electrochemical (EC) approaches that may have a chance to significantly improve an LFA’s performance that is needed in order to detect analytes reliably at lower concentrations than currently possible. In fact, EC-LFAs offer advantages in terms of quantitative determination, low-cost, high sensitivity, and even simple, label-free strategies. Here, the various configurations of EC-LFAs published are summarized and critically evaluated. In short, most of them rely on applying conventional transducers, e.g., screen-printed electrode, to ensure reliability of the assay, and additional advances are afforded by the beneficial features of nanomaterials. It is predicted that these will be further implemented in EC-LFAs as high-performance transducers. Considering the low cost of point-of-care devices, it becomes even more important to also identify strategies that efficiently integrate nanomaterials into EC-LFAs in a high-throughput manner while maintaining their favorable analytical performance.


Micromachines ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 697
Author(s):  
Siming Lu ◽  
Sha Lin ◽  
Hongrui Zhang ◽  
Liguo Liang ◽  
Shien Shen

Respiratory viral infections threaten human life and inflict an enormous healthcare burden worldwide. Frequent monitoring of viral antibodies and viral load can effectively help to control the spread of the virus and make timely interventions. However, current methods for detecting viral load require dedicated personnel and are time-consuming. Additionally, COVID-19 detection is generally relied on an automated PCR analyzer, which is highly instrument-dependent and expensive. As such, emerging technologies in the development of respiratory viral load assays for point-of-care (POC) testing are urgently needed for viral screening. Recent advances in loop-mediated isothermal amplification (LAMP), biosensors, nanotechnology-based paper strips and microfluidics offer new strategies to develop a rapid, low-cost, and user-friendly respiratory viral monitoring platform. In this review, we summarized the traditional methods in respiratory virus detection and present the state-of-art technologies in the monitoring of respiratory virus at POC.


Sensors ◽  
2021 ◽  
Vol 21 (3) ◽  
pp. 942
Author(s):  
Razvan Pascu ◽  
Gheorghe Pristavu ◽  
Gheorghe Brezeanu ◽  
Florin Draghici ◽  
Philippe Godignon ◽  
...  

A SiC Schottky dual-diode temperature-sensing element, suitable for both complementary variation of VF with absolute temperature (CTAT) and differential proportional to absolute temperature (PTAT) sensors, is demonstrated over 60–700 K, currently the widest range reported. The structure’s layout places the two identical diodes in close, symmetrical proximity. A stable and high-barrier Schottky contact based on Ni, annealed at 750 °C, is used. XRD analysis evinced the even distribution of Ni2Si over the entire Schottky contact area. Forward measurements in the 60–700 K range indicate nearly identical characteristics for the dual-diodes, with only minor inhomogeneity. Our parallel diode (p-diode) model is used to parameterize experimental curves and evaluate sensing performances over this far-reaching domain. High sensitivity, upwards of 2.32 mV/K, is obtained, with satisfactory linearity (R2 reaching 99.80%) for the CTAT sensor, even down to 60 K. The PTAT differential version boasts increased linearity, up to 99.95%. The lower sensitivity is, in this case, compensated by using a high-performing, low-cost readout circuit, leading to a peak 14.91 mV/K, without influencing linearity.


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