Ultrasensitive electrochemical biosensor based on <italic>in-situ</italic> labeling of electroactive melamine-Cu<sup>2+</sup> complex on the free terminal of the molecular beacon

2014 ◽  
Vol 44 (3) ◽  
pp. 381-388
Author(s):  
Le HANG ◽  
Feng GAO ◽  
Xuan ZHANG ◽  
QingXiang WANG ◽  
XiaoLei LIAO ◽  
...  
Keyword(s):  
Molecular Beacon ◽  
Electrochemical Biosensor

scholarly journals Single Step In Situ Detection of Surface Protein and MicroRNA in Clustered Extracellular Vesicles Using Flow Cytometry

2021 ◽  
Vol 10 (2) ◽  
pp. 319
Author(s):  
Hee Cheol Yang ◽  
Won Jong Rhee
Keyword(s):  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Molecular Beacon ◽  
Disease Diagnosis ◽  
Single Step ◽  
Flow Cytometer ◽  
Surface Proteins ◽  
Biomarker Detection ◽  
In Situ Detection

Because cancers are heterogeneous, it is evident that multiplexed detection is required to achieve disease diagnosis with high accuracy and specificity. Extracellular vesicles (EVs) have been a subject of great interest as sources of novel biomarkers for cancer liquid biopsy. However, EVs are nano-sized particles that are difficult to handle; thus, it is necessary to develop a method that enables efficient and straightforward EV biomarker detection. In the present study, we developed a method for single step in situ detection of EV surface proteins and inner miRNAs simultaneously using a flow cytometer. CD63 antibody and molecular beacon-21 were investigated for multiplexed biomarker detection in normal and cancer EVs. A phospholipid-polymer-phospholipid conjugate was introduced to induce clustering of the EVs analyzed using nanoparticle tracking analysis, which enhanced the detection signals. As a result, the method could detect and distinguish cancer cell-derived EVs using a flow cytometer. Thus, single step in situ detection of multiple EV biomarkers using a flow cytometer can be applied as a simple, labor- and time-saving, non-invasive liquid biopsy for the diagnosis of various diseases, including cancer.

Antidiabetic Drug Metformin Oxidation and in situ Interaction with dsDNA Using a dsDNA‐electrochemical Biosensor

2019 ◽  
Vol 31 (10) ◽  
pp. 1977-1987 ◽  
Author(s):  
W. B. S. Machini ◽  
I. P. G. Fernandes ◽  
A. M. Oliveira‐Brett
Keyword(s):  
Electrochemical Biosensor ◽  
Antidiabetic Drug

In Situ Evaluation of the Anticancer Antibody Rituximab-dsDNA Interaction Using a DNA-Electrochemical Biosensor

2014 ◽  
Vol 26 (6) ◽  
pp. 1304-1311 ◽  
Author(s):  
Inês B. Santarino ◽  
Severino Carlos B. Oliveira ◽  
Ana Maria Oliveira-Brett
Keyword(s):  
Electrochemical Biosensor ◽  
Dna Electrochemical Biosensor

scholarly journals Super-resolution imaging of a 2.5 kb non-repetitive DNA in situ in the nuclear genome using molecular beacon probes

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Yanxiang Ni ◽  
Bo Cao ◽  
Tszshan Ma ◽  
Gang Niu ◽  
Yingdong Huo ◽  
...  
Keyword(s):  
Repetitive Dna ◽  
Molecular Beacon ◽  
Genomic Sequence ◽  
Single Cells ◽  
Nuclear Genome ◽  
Super Resolution ◽  
Resolution Limit ◽  
Fish Method ◽  
Resolution Imaging

High-resolution visualization of short non-repetitive DNA in situ in the nuclear genome is essential for studying looping interactions and chromatin organization in single cells. Recent advances in fluorescence in situ hybridization (FISH) using Oligopaint probes have enabled super-resolution imaging of genomic domains with a resolution limit of 4.9 kb. To target shorter elements, we developed a simple FISH method that uses molecular beacon (MB) probes to facilitate the probe-target binding, while minimizing non-specific fluorescence. We used three-dimensional stochastic optical reconstruction microscopy (3D-STORM) with optimized imaging conditions to efficiently distinguish sparsely distributed Alexa-647 from background cellular autofluorescence. Utilizing 3D-STORM and only 29–34 individual MB probes, we observed 3D fine-scale nanostructures of 2.5 kb integrated or endogenous unique DNA in situ in human or mouse genome, respectively. We demonstrated our MB-based FISH method was capable of visualizing the so far shortest non-repetitive genomic sequence in 3D at super-resolution.

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