FRACTAL: Signal amplification of immunofluorescence via cyclic staining of target molecules

Nanoscale ◽  
2020 ◽  
Vol 12 (46) ◽  
pp. 23506-23513
Author(s):  
Yehlin Cho ◽  
Junyoung Seo ◽  
Yeonbo Sim ◽  
Jinkyoung Chung ◽  
Chan E. Park ◽  
...  
Keyword(s):  
Signal Intensity ◽  
Signal Amplification ◽  
Immunofluorescence Staining ◽  
Fractal Signal ◽  
Amplification Technique ◽  
Target Molecules ◽  
Secondary Antibodies

We demonstrate a novel signal amplification technique that can amplify the signal intensity of immunofluorescence staining via simple cyclic staining of secondary antibodies.

Sensitive detection of microRNA using QCM biosensors: sandwich hybridization and signal amplification by TiO2 nanoparticles

2020 ◽  
Vol 12 (42) ◽  
pp. 5103-5109
Author(s):  
Ji Yoon Lim ◽  
Soo Suk Lee
Keyword(s):  
Tio2 Nanoparticles ◽  
Signal Amplification ◽  
Sensitive Detection ◽  
Sandwich Hybridization ◽  
Amplification Technique

A QCM biosensor for the detection of miR-21 has been demonstrated along with sandwich hybridization and TiO2 nanoparticle-based photocatalytic signal amplification technique.

scholarly journals Evaluation of direct grafting strategies in Expansion Microscopy

2019 ◽  
Author(s):  
Gang Wen ◽  
Marisa Vanheusden ◽  
Aline Acke ◽  
Donato Vali ◽  
Simon Finn Mayer ◽  
...  
Keyword(s):  
Fine Structure ◽  
Lipid Membranes ◽  
Signal Amplification ◽  
Polymerization Reaction ◽  
Nanometer Scale ◽  
Biomolecular Systems ◽  
Covalent Grafting ◽  
Two Factors ◽  
Secondary Antibodies ◽  
Polymerizable Group

AbstractHigh resolution fluorescence microscopy is a key tool in the elucidation of biological fine-structure, providing insights into the distribution and interactions of biomolecular systems down to the nanometer scale. Expansion microscopy is a recently developed approach to achieving nanoscale resolution in optical imaging. In the experiment, biological samples are embedded in a hydrogel, which is isotropicaly swollen. This physically pulls labels apart, allowing more of them to be resolved. However, in the gelation and swelling process, two factors combine to reduce the signal in the final image; signal dilution and the polymerization reaction, which can damage some fluorophores. Here, we show a chemical linking approach that allows covalent grafting of biomolecular target and reporter in expansion microscopy. Through the combination of a targeting ligand, a reporter moiety and a polymerizable group in a single linker, complex constructs can be prepared in a single, labelling step. We show application of this new series of molecules in the targeting of the cell cytoskeleton, a first example of lipid membranes in expansion microscopy; direct immunostaining with primary and secondary antibodies, and direct grafting of ISH probes and signal amplification initiators (HCR and RollFISH). Our probes allow direct, multiplexed targeting of the cellular blueprint and enable a range of novel imaging approaches in combination with expansion microscopy.

Detection of thrombin based on aptamer isothermal exponential signal amplification technique

The Analyst ◽  
2015 ◽  
Vol 140 (19) ◽  
pp. 6489-6492 ◽  
Author(s):  
Xiaotong Shen ◽  
Menghua Zhang ◽  
Shuyan Niu ◽  
Chao Shi
Keyword(s):  
Signal Amplification ◽  
Protein Detection ◽  
Amplification Technique

In this report, a strategy based on an aptameric molecule switch to realize homogeneous and isothermal signal exponential amplification and protein detection has been described.

Aptamer-Based Sensitive Detection of Target Molecules via RT-PCR Signal Amplification

2010 ◽  
Vol 21 (12) ◽  
pp. 2183-2189 ◽  
Author(s):  
Shiqi Liao ◽  
Yuanqiang Liu ◽  
Jiayu Zeng ◽  
Xiaoxu Li ◽  
Ningsheng Shao ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Sensitive Detection ◽  
Rt Pcr ◽  
Target Molecules

Enzyme-assisted target recycling (EATR) for nucleic acid detection

2014 ◽  
Vol 43 (17) ◽  
pp. 6405-6438 ◽  
Author(s):  
Yulia V. Gerasimova ◽  
Dmitry M. Kolpashchikov
Keyword(s):  
Nucleic Acid ◽  
Signal Amplification ◽  
Nucleic Acid Detection ◽  
Target Recycling ◽  
Amplification Technique

Enzyme-assisted target recycling (EATR) is a signal amplification technique that can find application in PCR-free nucleic acid detection.

scholarly journals An Enzyme-Free Signal Amplification Technique for Ultrasensitive Colorimetric Assay of Disease Biomarkers

ACS Nano ◽  
2017 ◽  
Vol 11 (2) ◽  
pp. 2052-2059 ◽  
Author(s):  
Haihang Ye ◽  
Kuikun Yang ◽  
Jing Tao ◽  
Yijing Liu ◽  
Qian Zhang ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Colorimetric Assay ◽  
Disease Biomarkers ◽  
Amplification Technique

scholarly journals A multiplex microchamber diffusion assay for the antibody-based detection of microRNAs on randomly ordered microbeads

2021 ◽  
Author(s):  
Christiane Geithe ◽  
Bo Zeng ◽  
Carsten Schmidt ◽  
Franziska Dinter ◽  
Dirk Roggenbuck ◽  
...  
Keyword(s):  
Gene Expression ◽  
Diffusion Process ◽  
Regulation Of Gene Expression ◽  
P Value ◽  
Clinical Biomarkers ◽  
Microscope Imaging ◽  
Target Molecules ◽  
Mirna Detection ◽  
Secondary Antibodies ◽  
Post Transcriptional Regulation

Background: MicroRNAs (miRNAs) are small, conserved, noncoding RNAs regulating gene expression that functions in RNA silencing and post-transcriptional regulation of gene expression. Altered miRNA profiles have been implicated in many human diseases, and due to their circulating abilities, they have excited great interest in their use as clinical biomarkers. The development of innovative methods for miRNA detection has become of high scientific and clinical interest. Methods: We developed a diffusion-driven microbead assay and combined it with an antibody-based miRNA detection. The diffusion process was carried out in two different approaches a) co-diffusion of miRNA and antibodies (termed diffusion approach I, DAI) and b) diffusion of miRNA in an antibody-saturated environment (DAII). In both approaches, neutravidin-coated microbeads were loaded with specific biotinylated DNA capture probes, which targets either miR-21-5p, miR-30a-3p or miR-93-5p. The miRNAs were time- and dose-dependently detected in a diffusion microchamber by primary anti-DNA:RNA hybrid and fluorescence-labeled secondary antibodies using our in-house developed inverse fluorescence microscope imaging platform VideoScan. Results: Our assay offers the advantage that several target molecules can be detected simultaneously and in real-time in one reaction environment (multiplex), without any amplification steps. We recorded the diffusion process over a period of 24 h and found that the reaction was almost completed after 2 h. The specificity of the assay was 96.7 % for DAI and 92.3 % for DAII. The detection limits were in a concentration range of 0.03-0.43 nM for DAI and 0.14-1.09 nM for DAII, depending on the miRNA. Conclusion: The miRNAs are successively exposed to the capture probe-loaded randomly ordered microbeads (p value of CSR 0.23-0.96), which leads to microbeads that become saturated with the target molecules first in front rows. Non-bonded miRNAs continue to diffuse further and can therefore subsequently bind to the microbeads with free binding sites. Our detection principle differs from other microbead assays, in which all microbeads are simultaneously mixed with the sample solution, so that all target molecules bind equally distributed to the microbeads, resulting in an averaged signal intensity.

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