scholarly journals Quantifying Heterogeneity According to Deformation of the U937 Monocytes and U937-Differentiated Macrophages Using 3D Carbon Dielectrophoresis in Microfluidics

Micromachines ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 576 ◽  
Author(s):  
Meltem Elitas ◽  
Esra Sengul
Keyword(s):  
Fluid Shear Stress ◽  
Cell Damage ◽  
Electrode Array ◽  
Crossover Frequency ◽  
Label Free ◽  
Electromechanical Properties ◽  
Maximum Deformation ◽  
Deformation Index ◽  
Minimum Deformation ◽  
Downstream Analysis

A variety of force fields have thus far been demonstrated to investigate electromechanical properties of cells in a microfluidic platform which, however, are mostly based on fluid shear stress and may potentially cause irreversible cell damage. This work presents dielectric movement and deformation measurements of U937 monocytes and U937-differentiated macrophages in a low conductive medium inside a 3D carbon electrode array. Here, monocytes exhibited a crossover frequency around 150 kHz and presented maximum deformation index at 400 kHz and minimum deformation index at 1 MHz frequencies at 20 Vpeak-peak. Although macrophages were differentiated from monocytes, their crossover frequency was lower than 50 kHz at 10 Vpeak-peak. The change of the deformation index for macrophages was more constant and lower than the monocyte cells. Both dielectric mobility and deformation spectra revealed significant differences between the dielectric responses of U937 monocytes and U937-differentiated macrophages, which share the same origin. This method can be used for label-free, specific, and sensitive single-cell characterization. Besides, damage of the cells by aggressive shear forces can, hence, be eliminated and cells can be used for downstream analysis. Our results showed that dielectric mobility and deformation have a great potential as an electromechanical biomarker to reliably characterize and distinguish differentiated cell populations from their progenitors.

scholarly journals Label-free prediction of Cell Painting from brightfield images

2021 ◽  
Author(s):  
Jan Oscar Cross-Zamirski ◽  
Elizabeth Mouchet ◽  
Guy Williams ◽  
Carola-Bibiane Schönlieb ◽  
Riku Turkki ◽  
...  
Keyword(s):  
Pearson Correlation ◽  
Ground Truth ◽  
Free Cell ◽  
Label Free ◽  
Biological Interest ◽  
Genetic Perturbations ◽  
Toxicity Analysis ◽  
Model Training ◽  
Downstream Analysis ◽  
Cellular Components

Cell Painting is a high-content image-based assay which can reveal rich cellular morphology and is applied in drug discovery to predict bioactivity, assess toxicity and understand diverse mechanisms of action of chemical and genetic perturbations. In this study, we investigate label-free Cell Painting by predicting the five fluorescent Cell Painting channels from paired brightfield z-stacks using deep learning models. We train and validate the models with a dataset representing 1000s of pan-assay interference compounds sampled from 17 unique batches. The model predictions are evaluated using a test set from two additional batches, treated with compounds comprised from a publicly available phenotypic set. In addition to pixel-level evaluation, we process the label-free Cell Painting images with a segmentation-based feature-extraction pipeline to understand whether the generated images are useful in downstream analysis. The mean Pearson correlation coefficient (PCC) of the images across all five channels is 0.84. Without actually incorporating these features into the model training we achieved a mean correlation of 0.45 from the features extracted from the images. Additionally we identified 30 features which correlated greater than 0.8 to the ground truth. Toxicity analysis on the label-free Cell Painting resulted a sensitivity of 62.5% and specificity of 99.3% on images from unseen batches. Additionally, we provide a breakdown of the feature profiles by channel and feature type to understand the potential and limitation of the approach in morphological profiling. Our findings demonstrate that label-free Cell Painting has potential above the improved visualization of cellular components, and it can be used for downstream analysis. The findings also suggest that label-free Cell Painting could allow for repurposing the imaging channels for other non-generic fluorescent stains of more targeted biological interest, thus increasing the information content of the assay.

scholarly journals Accuracy analysis and optimization of infrared guidance test device

2020 ◽  
Vol 12 (6) ◽  
pp. 168781402092265
Author(s):  
Zhou Wang ◽  
Yin Chen ◽  
Tao Wang ◽  
Bo Zhang
Keyword(s):  
Physical Simulation ◽  
Orthogonal Experiment ◽  
Target Position ◽  
Design Parameters ◽  
Test Device ◽  
Factors Affecting ◽  
Pointing Error ◽  
Maximum Deformation ◽  
Minimum Deformation ◽  
Shell Optimization

As an important modern weapon, the development of infrared-guided missile reflects comprehensive national strength of a country. Therefore, it is especially important to establish a semi-physical simulation device to test the performance of missile, and the test device requires high accuracy. Based on the above background, an infrared guidance test device is designed in this article. The accuracy of its shell and rotating mechanism are studied in detail, and the error factors are quantified to provide theoretical basis for structural optimization. The orthogonal experiment design reduces the number of sensitivity analysis experiments on key design parameters. Factors affecting the maximum deformation and overall quality of the shell were determined. The range method was used to analyze sensitivity factors, and the final optimization result that met the minimum deformation and minimum quality was determined. Experimental results show that the rotation error of the main shaft of the rotating mechanism includes axial, radial, and angular motion errors, and experimental value is basically consistent with theoretical value. After the shell optimization, the infrared target pointing error [Formula: see text] and the infrared target position offset error ξ′ = 0.1525 mm meet the accuracy requirements. This method can provide new ideas for precision research and optimization of structural design of rotating mechanism.

Effect of Electrical Pulsing on Various Heat Treatments of 5XXX Series Aluminum Alloys

2008 ◽  
Author(s):  
Wesley A. Salandro ◽  
Joshua J. Jones ◽  
Timothy A. McNeal ◽  
John T. Roth ◽  
Sung-Tae Hong ◽  
...  
Keyword(s):  
Aluminum Alloys ◽  
Superplastic Deformation ◽  
Deformation Process ◽  
Electrical Current ◽  
Heat Treatments ◽  
Maximum Deformation ◽  
Aluminum Specimen ◽  
Electrical Pulses ◽  
Minimum Deformation ◽  
Diffuse Neck

Previous studies have shown that the presence of a pulsed electrical current, applied during the deformation process of an aluminum specimen, can significantly improve the formability of the aluminum without heating the metal above its maximum operating temperature range. The research herein extends these findings by examining the effect of electrical pulsing on 5052 and 5083 Aluminum Alloys. Two different parameter sets were used while pulsing three different heat treatments (As Is, 398°C, and 510°C) for each of the two aluminum alloys. For this research, the electrical pulsing is applied to the aluminum while the specimens are deformed, without halting the deformation process. The analysis focuses on establishing the effect the electrical pulsing has on the aluminum alloy’s various heat treatments by examining the displacement of the material throughout the testing region of dogbone shaped specimens. The results from this research show that pulsing significantly increases the maximum achievable elongation of the aluminum (when compared to baseline tests conducted without electrical pulsing). Significantly reducing the engineering flow stress within the material is another beneficial effect produced by electric pulsing. The electrical pulses also cause the aluminum to deform non-uniformly, such that the material exhibits a diffuse neck where the minimum deformation occurs near the ends of the specimen (near the clamps) and the maximum deformation occurs near the center of the specimen (where fracture ultimately occurs). This diffuse necking effect is similar to what can be experienced during superplastic deformation. However, when comparing the presence of a diffuse neck in this research, electrical pulsing does not create as significant of a diffuse neck as superplastic deformation. Electrical pulsing has the potential to be more efficient than traditional methods of incremental forming since the deformation process is never interrupted. Overall, with the greater elongation and lower stress, the aluminum can be deformed quicker, easier, and to a greater extent than is currently possible.

scholarly journals Immuno-Acoustic Sorting of Disease-Specific Extracellular Vesicles by Acoustophoretic Force

Micromachines ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1534
Author(s):  
Junyuan Liu ◽  
Yuxin Qu ◽  
Han Wang
Keyword(s):  
Extracellular Vesicles ◽  
Processing Time ◽  
Life Science ◽  
Biological Fluids ◽  
Label Free ◽  
Her2 Positive ◽  
Diagnosis And Prognosis ◽  
Efficient Recovery ◽  
Downstream Analysis ◽  
Disease Specific

Methods for the isolation and analysis of extracellular vesicles (EVs) have been extensively explored in the field of life science and in clinical diagnosis in recent years. The separation and efficient recovery of high-purity target EVs from biological samples are important prerequisites in the study of EVs. So far, commonly used methods of EV separation include ultracentrifugation, filtration, solvent precipitation and immunoaffinity capturing. However, these methods suffer from long processing time, EV damage and low enrichment efficiency. The use of acoustophoretic force facilitates the non-contact label-free manipulation of cells based on their size and compressibility but lacks specificity. Additionally, the acoustophoretic force exerted on sub-micron substances is normally weak and insufficient for separation. Here we present a novel immuno-acoustic sorting technology, where biological substances such as EVs, viruses, and biomolecules, can be specifically captured by antibody/receptor coated microparticles through immunoaffinity, and manipulated by an acoustophoretic force exerted on the microparticles. Using immuno-acoustic sorting technology, we successfully separated and purified HER2-positive EVs for further downstream analysis. This method holds great potential in isolating and purifying specific targets such as disease-related EVs from biological fluids and opens new possibilities for the EV-based early diagnosis and prognosis of diseases.

scholarly journals Robust summarization and inference in proteome-wide label-free quantification

2019 ◽  
Author(s):  
Adriaan Sticker ◽  
Ludger Goeminne ◽  
Lennart Martens ◽  
Lieven Clement
Keyword(s):  
Data Analysis ◽  
State Of The Art ◽  
Downstream Processing ◽  
Superior Performance ◽  
Model Complexity ◽  
Model Parameters ◽  
Label Free ◽  
Specific Effects ◽  
Modular Analysis ◽  
Downstream Analysis

AbstractLabel-Free Quantitative mass spectrometry based workflows for differential expression (DE) analysis of proteins impose important challenges on the data analysis due to peptide-specific effects and context dependent missingness of peptide intensities. Peptide-based workflows, like MSqRob, test for DE directly from peptide intensities and outper-form summarization methods which first aggregate MS1 peptide intensities to protein intensities before DE analysis. However, these methods are computationally expensive, often hard to understand for the non-specialised end-user, and do not provide protein summaries, which are important for visualisation or downstream processing. In this work, we therefore evaluate state-of-the-art summarization strategies using a benchmark spike-in dataset and discuss why and when these fail compared to the state-of-the-art peptide based model, MSqRob. Based on this evaluation, we propose a novel summarization strategy, MSqRob-Sum, which estimates MSqRob’s model parameters in a two-stage procedure circumventing the drawbacks of peptide-based workflows. MSqRobSum maintains MSqRob’s superior performance, while providing useful protein expression summaries for plotting and downstream analysis. Summarising peptide to protein intensities considerably reduces the computational complexity, the memory footprint and the model complexity, and makes it easier to disseminate DE inferred on protein summaries. Moreover, MSqRobSum provides a highly modular analysis framework, which provides researchers with full flexibility to develop data analysis workflows tailored towards their specific applications.

Formability of Al 5xxx Sheet Metals Using Pulsed Current for Various Heat Treatments

2010 ◽  
Vol 132 (5) ◽  
Author(s):  
Wesley A. Salandro ◽  
Joshua J. Jones ◽  
Timothy A. McNeal ◽  
John T. Roth ◽  
Sung-Tae Hong ◽  
...  
Keyword(s):  
Aluminum Alloys ◽  
Superplastic Deformation ◽  
Deformation Process ◽  
Electrical Current ◽  
Heat Treatments ◽  
Maximum Deformation ◽  
Aluminum Specimen ◽  
Electrical Pulses ◽  
Minimum Deformation ◽  
Diffuse Neck

Previous studies have shown that the presence of a pulsed electrical current, applied during the deformation process of an aluminum specimen, can significantly improve the formability of the aluminum without heating the metal above its maximum operating temperature range. The research herein extends these findings by examining the effect of electrical pulsing on 5052 and 5083 aluminum alloys. Two different parameter sets were used while pulsing three different heat-treatments (as-is, 398°C, and 510°C) for each of the two aluminum alloys. For this research, the electrical pulsing is applied to the aluminum while the specimens are deformed without halting the deformation process (a manufacturing technique known as electrically assisted manufacturing). The analysis focuses on establishing the effect of the electrical pulsing has on the aluminum alloy’s various heat-treatments by examining the displacement of the material throughout the testing region of dogbone-shaped specimens. The results from this research show that pulsing significantly increases the maximum achievable elongation of the aluminum (when compared with baseline tests conducted without electrical pulsing). Another beneficial effect produced by electrical pulsing is that the engineering flow stress within the material is considerably reduced. The electrical pulses also cause the aluminum to deform nonuniformly, such that the material exhibits a diffuse neck where the minimum deformation occurs near the ends of the specimen (near the clamps) and the maximum deformation occurs near the center of the specimen (where fracture ultimately occurs). This diffuse necking effect is similar to what can be experienced during superplastic deformation. However, when comparing the presence of a diffuse neck in this research, electrical pulsing does not create as significant of a diffuse neck as superplastic deformation. Electrical pulsing has the potential to be more efficient than the traditional methods of incremental forming since the deformation process is never interrupted. Overall, with the greater elongation and lower stress, the aluminum can be deformed quicker, easier, and to a greater extent than is currently possible.

scholarly journals Label-Free Impedance Sensing of Aflatoxin B1 with Polyaniline Nanofibers/Au Nanoparticle Electrode Array

Sensors ◽  
2018 ◽  
Vol 18 (5) ◽  
pp. 1320 ◽  
Author(s):  
Ajay Kumar Yagati ◽  
Sachin Ganpat Chavan ◽  
Changyoon Baek ◽  
Min-Ho Lee ◽  
Junhong Min
Keyword(s):  
Aflatoxin B1 ◽  
Electrode Array ◽  
Au Nanoparticle ◽  
Label Free ◽  
Impedance Sensing ◽  
Polyaniline Nanofibers

scholarly journals Digital Staining of High-Definition Fourier Transform Infrared (FT-IR) Images Using Deep Learning

2019 ◽  
Vol 73 (5) ◽  
pp. 556-564 ◽  
Author(s):  
Mahsa Lotfollahi ◽  
Sebastian Berisha ◽  
Davar Daeinejad ◽  
David Mayerich
Keyword(s):  
Fourier Transform ◽  
Fourier Transform Infrared ◽  
Cellular Level ◽  
Label Free ◽  
High Definition ◽  
Tissue Samples ◽  
Ft Ir ◽  
Downstream Analysis ◽  
Ir Spectroscopic ◽  
Multiple Samples

Histological stains, such as hematoxylin and eosin (H&E), are routinely used in clinical diagnosis and research. While these labels offer a high degree of specificity, throughput is limited by the need for multiple samples. Traditional histology stains, such as immunohistochemical labels, also rely only on protein expression and cannot quantify small molecules and metabolites that may aid in diagnosis. Finally, chemical stains and dyes permanently alter the tissue, making downstream analysis impossible. Fourier transform infrared (FT-IR) spectroscopic imaging has shown promise for label-free characterization of important tissue phenotypes and can bypass the need for many chemical labels. Fourier transform infrared classification commonly leverages supervised learning, requiring human annotation that is tedious and prone to errors. One alternative is digital staining, which leverages machine learning to map IR spectra to a corresponding chemical stain. This replaces human annotation with computer-aided alignment. Previous work relies on alignment of adjacent serial tissue sections. Since the tissue samples are not identical at the cellular level, this technique cannot be applied to high-definition FT-IR images. In this paper, we demonstrate that cellular-level mapping can be accomplished using identical samples for both FT-IR and chemical labels. In addition, higher-resolution results can be achieved using a deep convolutional neural network that integrates spatial and spectral features.

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