scholarly journals Integrated Field’s metal microelectrodes based microfluidic impedance cytometry for cell-in-droplet quantification

2018 ◽  
Author(s):  
Jatin Panwar ◽  
Rahul Roy
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Microfluidic Channel ◽  
High Sensitivity ◽  
Cell Detection ◽  
Label Free ◽  
Single Cell Detection ◽  
Electrode Fabrication ◽  
Impedance Cytometry

AbstractMicrofluidic impedance cytometry (MIC) provides a non-optical and label-free method for single cell detection and classification in microfluidics. However, the cleanroom intensive infrastructure required for MIC electrode fabrication limits its wide implementation in microfluidic analysis. To bypass the conventional metal (platinum) electrode fabrication protocol, we fabricated coplanar ‘in-contact’ Field’s metal (icFM) microelectrodes in multilayer elastomer devices with a single photolithography step. Our icFM microelectrodes displayed excellent and comparable performance to the platinum electrodes for detection of single erythrocytes with a lock-in amplifier based MIC setup. We further characterized it for water-in-oil droplets generated in a T-junction microfluidic channel and found high sensitivity and long-term operational stability of these electrodes. Finally, to facilitate droplet based single cell analysis, we demonstrate detection and quantification of single cells entrapped in aqueous droplets.

scholarly journals Multiplexed and single-cell detection of microRNA with plasmonic nanoparticle assemblies

2021 ◽  
Author(s):  
Gurbrinder Ghotra ◽  
Nguyen Le ◽  
Heyam Hayder ◽  
Chun Peng ◽  
Jennifer I. L. Chen
Keyword(s):  
Nucleic Acid ◽  
Single Cell ◽  
Low Cost ◽  
Single Cells ◽  
Cell Detection ◽  
Label Free ◽  
Plasmonic Nanoparticle ◽  
Nanoparticle Assemblies ◽  
Single Cell Detection ◽  
Target Binding

We present a label-free, low cost and miniatured biosensing platform based on the disassembly of core-satellite plasmonic nanoparticle assemblies. The rapid and selective detection of an exemplary nucleic acid biomarker, has-miRNA-210-3p, was achieved via the strand displacement nucleic acid reaction. Target binding leads to dehybridization of the DNA linkers and changes in the scattering properties of nanostructures as monitored by darkfield microscopy. We demonstrate the ability to detect microRNA expunged from single cells and the potential to multiplex discrete assemblies to enable diverse biological applicability. The work may help translate the applicability of microRNA as diagnostic biomarkers, quantitate their abundance in the microenvironment, and facilitate the study of their correlation or causation to other biomolecules at the single-cell level.

scholarly journals Single-cell detection of Nonsense-Mediated mRNA decay

2021 ◽  
Author(s):  
Robert Singer ◽  
Hanae Sato
Keyword(s):  
Single Cell ◽  
Mrna Decay ◽  
Single Cells ◽  
Mrna Degradation ◽  
Cell Detection ◽  
Globin Genes ◽  
Reporter System ◽  
Single Cell Detection ◽  
Translational Readthrough ◽  
Nonsense Mediated Mrna Decay

Abstract Nonsense-mediated mRNA decay (NMD) is an mRNA degradation pathway that eliminates transcripts containing premature termination codons (PTCs). Half-lives of the mRNAs containing PTCs demonstrates that a small percent escape surveillance and do not degrade. It is not known whether this escape represents variable mRNA degradation within cells or, alternatively cells within the population are resistant. Here we demonstrate a single-cell approach with a bi-directional reporter, which expresses two b-globin genes with or without a PTC in the same cell, to characterize the efficiency of NMD in individual cells. We found a broad range of NMD efficiency in the population; some cells degraded essentially all of the mRNAs, while others escaped NMD almost completely. Characterization of NMD efficiency together with NMD regulators in single cells showed cell-to-cell variability of NMD reflects the differential level of surveillance factors, SMG1 and phosphorylated UPF1. A single-cell fluorescent reporter system that enabled detection of NMD using flow cytometry revealed that this escape occurred either by translational readthrough at the PTC or by failure of mRNA degradation after successful translation termination at the PTC.

A sheath-less electric impedance micro-flow cytometry device for rapid label-free cell classification and viability testing

2017 ◽  
Vol 9 (7) ◽  
pp. 1201-1212 ◽  
Author(s):  
Xinwu Xie ◽  
Zhen Cheng ◽  
Youchun Xu ◽  
Ran Liu ◽  
Qi Li ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Simple Structure ◽  
Electrical Conductance ◽  
Free Cell ◽  
Cell Detection ◽  
Label Free ◽  
Electric Impedance ◽  
Single Cell Detection ◽  
Micro Flow

A sheath-less PDMS microfluidic IFC device with a simple structure was constructed, with a good performance in single-cell detection. The electrical conductance and susceptance were used to differentiate the beads/cells.

scholarly journals High-throughput label-free molecular fingerprinting flow cytometry

2019 ◽  
Vol 5 (1) ◽  
pp. eaau0241 ◽  
Author(s):  
Kotaro Hiramatsu ◽  
Takuro Ideguchi ◽  
Yusuke Yonamine ◽  
SangWook Lee ◽  
Yizhi Luo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Fluorescent Labeling ◽  
Live Cells ◽  
Molecular Fingerprinting ◽  
Label Free ◽  
Indispensable Tool

Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm−1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics ofHaematococcus lacustris.

Microfluidics Design for Single Cell Detection

2013 ◽  
Vol 562-565 ◽  
pp. 589-593
Author(s):  
Shao Bo Du ◽  
Sheng Bo Sang ◽  
Wen Dong Zhang ◽  
Jie Hu ◽  
Peng Wei Li ◽  
...  
Keyword(s):  
Single Cell ◽  
Fluid Transport ◽  
Individual Cell ◽  
Single Cells ◽  
Cell Detection ◽  
Cell Capture ◽  
Single Cell Detection ◽  
Novel Structure ◽  
Mother And Daughter

Here we demonstrate a microfluidic-based analysis system based on single cell capture array, which can physically trap individual cell using micrometer-sized structures. A stable and in vivo-like microenvironment was built with the novel structure at the single-cell detection level. The microfluidic-based design can decouple single cells from fluid flow with the help of micropillars. The size and geometry of the cell jails are designed in order to discriminate between mother and daughter cells. It provides an experimental platform to efficiently monitor individual cell state for a long period of time. Furthermore, the parallel microfluidic array can ensure accuracy. In addition, finite element method (FEM) was employed to predict fluid transport properties for the most optimal fluid microenvironment.

CMOS Biosensor IC with 360-Sensing Elements Using 63-GHz LC-Oscillator and DEP for Label-Free Single-Cell Detection

2018 ◽  
Author(s):  
Takeshi Mitsunaka ◽  
Masafumi Yamanoue ◽  
Kunihiko Iizuka ◽  
Soo Hyon Kim ◽  
Teruo Fujii ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Detection ◽  
Label Free ◽  
Single Cell Detection ◽  
Lc Oscillator

MULTISPECTRAL CONCURRENT DETECTION OF MULTIPLE PROTEINS

Nano LIFE ◽  
2012 ◽  
Vol 02 (03) ◽  
pp. 1241004 ◽  
Author(s):  
JINGWEN CHAI ◽  
QING SONG
Keyword(s):  
Single Cell ◽  
Measurement Errors ◽  
Single Cells ◽  
Cell Detection ◽  
Cell Level ◽  
Single Cell Detection ◽  
Real Time Measurement ◽  
Novel Approach ◽  
Clinical Diagnoses ◽  
Cell Sample

Proteins constitutively function within networks. Concurrent detection of multiple proteins is crucial to clinical diagnoses and multidimensional drug profiling. Fluorescence microscopy is capable of multicolor imaging, and has the capability to quantify essentially any physiological changes that occur at the single-cell level and in the context of live single cells, and thus provides an alternative to flow cytometry for multiplexed live single-cell assay. The staining of cells with multiple labels is still a technical challenge while multiplexed assays are complicated by spectral emission overlaps and measurement errors. In this study, we applied emission fingerprinting technique provided by Zeiss LSM 510 META detector, and achieved concurrent detection of ten proteins expressed on the same endothelial cell sample. This approach can be further applied to real-time measurement of multiple proteins expressed on live single cell surface, and therefore will enable a novel approach of multiplexed live single cell detection.

scholarly journals Multiplexed single-cell proteomics using SCoPE2

2021 ◽  
Author(s):  
Aleksandra A Petelski ◽  
Edward Emmott ◽  
Andrew Leduc ◽  
R. Gray Huffman ◽  
Harrison Specht ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Cost Effective ◽  
Protein Analysis ◽  
Single Cell Protein ◽  
Protein Quantification ◽  
Cell Protein ◽  
Label Free ◽  
Cost Effective Method

Many biological systems are composed of diverse single cells. This diversity necessitates functional and molecular single-cell analysis. Single-cell protein analysis has long relied on affinity reagents, but emerging mass-spectrometry methods (either label-free or multiplexed) have enabled quantifying over 1,000 proteins per cell while simultaneously increasing the specificity of protein quantification. Isobaric carrier based multiplexed single-cell proteomics is a scalable, reliable, and cost-effective method that can be fully automated and implemented on widely available equipment. It uses inexpensive reagents and is applicable to any sample that can be processed to a single-cell suspension. Here we describe an automated Single Cell ProtEomics (SCoPE2) workflow that allows analyzing about 200 single cells per 24 hours using only standard commercial equipment. We emphasize experimental steps and benchmarks required for achieving quantitative protein analysis.

A SIMULATION STUDY OF SINGLE CELL INSIDE AN INTEGRATED DUAL NANONEEDLE-MICROFLUDIC SYSTEM

2016 ◽  
Vol 78 (7-5) ◽  
Author(s):  
Muhammd Asraf Mansor ◽  
Mohd Ridzuan Ahmad
Keyword(s):  
Single Cell ◽  
Simulation Study ◽  
Single Cell Analysis ◽  
Biological Activities ◽  
Single Cells ◽  
Microfluidic Channel ◽  
Electrical Measurement ◽  
Solution Flow Rate ◽  
Solution Flow ◽  
Novel Method

Electrical properties of living cells have been proven to play significant roles in understanding of various biological activities including disease progression both at the cellular and molecular levels. Analyzing the cell’s electrical states especially in single cell analysis (SCA) lead to differentiate between normal cell and cancer cell. This paper presents a simulation study of micro-channel and nanoneedle structure, fluid manipulation and current flow through HeLa cell inside a microfluidic channel. To perform electrical measurement, gold dual nanoneedle has been utilized. The simulation result revealed, the cell penetration occurs at microchannel dimension and solution flow rate is 22 µm x 70 µm x 25 µm (width x length x height) and 0.396 pL/min, respectively. The purposed device has capability to characterize the electrical property of single cells can be used as a novel method for cell viability detection in instantaneous manner.

scholarly journals High throughput single-cell detection of multiplex CRISPR-edited gene modifications

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Elisa ten Hacken ◽  
Kendell Clement ◽  
Shuqiang Li ◽  
María Hernández-Sánchez ◽  
Robert Redd ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Single Cell ◽  
High Throughput ◽  
Gene Editing ◽  
Somatic Mutations ◽  
Single Cells ◽  
Lymphocytic Leukemia ◽  
Cell Detection ◽  
Loss Of Function ◽  
Single Cell Detection

Abstract CRISPR-Cas9 gene editing has transformed our ability to rapidly interrogate the functional impact of somatic mutations in human cancers. Droplet-based technology enables the analysis of Cas9-introduced gene edits in thousands of single cells. Using this technology, we analyze Ba/F3 cells engineered to express single or multiplexed loss-of-function mutations recurrent in chronic lymphocytic leukemia. Our approach reliably quantifies mutational co-occurrences, zygosity status, and the occurrence of Cas9 edits at single-cell resolution.

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