A ratiometric photoelectrochemical immunosensor based on g-C3N4@TiO2 NTs amplified by signal antibodies–Co3O4 nanoparticle conjugates

The Analyst ◽  
2018 ◽  
Vol 143 (20) ◽  
pp. 5030-5037 ◽  
Author(s):  
Qiong Wu ◽  
Fengxia Zhang ◽  
Huijuan Li ◽  
Zhihua Li ◽  
Qi Kang ◽  
...  
Keyword(s):  
Signal Amplification ◽  
Tio2 Nts ◽  
Secondary Antibodies

Herein, we report a ratiometric photoelectrochemical (PEC) immunosensor coupled with secondary antibodies–Co3O4 nanoparticle conjugates (Ab2–Co3O4 NPs) for signal amplification.

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.

scholarly journals Tyramide Signal Amplification coupled with multiple immunolabeling and RNAscope in situ hybridization in formaldehyde-fixed paraffin-embedded human fetal brain.

2021 ◽  
Author(s):  
Ayman Alzu'bi ◽  
Niveditha Sankar ◽  
Gavin Clowry
Keyword(s):  
Signal Amplification ◽  
Fetal Brain ◽  
Morphological Characteristics ◽  
High Sensitivity ◽  
Tyramide Signal Amplification ◽  
Human Fetal Brain ◽  
Immunofluorescence Labelling ◽  
Complex Detection ◽  
Secondary Antibodies

Abstract Several strategies have been recently introduced to improve the practicality of multiple immunolabelling and RNA in situ hybridization methods. We present a modified hybrid protocol of recently described complex detection strategies: (1) elution of antibodies prior to second round of staining (2) use of integrated polymers of HRP with secondary antibodies, and (3) tyramide signal amplification of multiple immunofluorescence labelling, to achieve a high sensitivity sequential multiple labeling using antibodies from the same species. A modified protocol of the novel RNAscope in situ hybridization method, including coupling with immunofluorescence on sections of early human fetal brain, has also been developed. These two techniques, when properly optimized, were highly compatible with routine formaldehyde-fixed paraffin-embedded tissue that preserves the best morphological characteristics of delicate fetal brain samples, allowing high power signal amplification for detection of protein and mRNA of genes that are sparsely expressed in the human fetal telencephalon.

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.

scholarly journals Indexable signal amplification for multiparametric imaging

2021 ◽  
Author(s):  
Paul D Simonson ◽  
Itzel Valencia ◽  
Sanjay S Patel
Keyword(s):  
Signal Amplification ◽  
Trial And Error ◽  
Dna Barcodes ◽  
Detailed Characterization ◽  
Multiparametric Imaging ◽  
Antibody Staining ◽  
Antibody Panel ◽  
Secondary Antibodies

Multiparametric imaging allows researchers to measure the expression of many biomarkers simultaneously, allowing detailed characterization of cell microenvironments. One such technique, CODEX, allows fluorescence imaging of >30 proteins in a single tissue section. In the commercial CODEX system, primary antibodies are conjugated to DNA barcodes. This modification can result in antibody dysfunction, and development of a custom antibody panel can be very costly and time consuming as trial and error of modified antibodies proceeds. To address these challenges, we developed novel tyramide-conjugated DNA barcodes that can be used with primary antibodies via peroxidase-conjugated secondary antibodies. This approach results in signal amplification and imaging without the need to conjugate primary antibodies. When combined with commercially available barcode-conjugated primary antibodies, we can very quickly develop working antibody panels. We also present methods to perform antibody staining using a commercially available automated tissue stainer and in situ hybridization imaging on a CODEX platform.

Visualization of von Willebrand Factor Multimers by Enzyme-Conjugated Secondary Antibodies

1986 ◽  
Vol 55 (02) ◽  
pp. 276-278 ◽  
Author(s):  
F Brosstad ◽  
Inge Kjønniksen ◽  
B Rønning ◽  
H Stormorken
Keyword(s):  
Von Willebrand Factor ◽  
Chromogenic Substrate ◽  
Agarose Electrophoresis ◽  
Secondary Antibodies ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Beta Galactosidase

SummaryA method for visualization of the multimeric forms of von Willebrand Factor (vWF) in plasma and platelets is described. The method is based upon: 1) Separation of the vWF multimers by SDS-agarose electrophoresis, 2) Subsequent blotting of the vWF multimers onto nitrocellulose, 3) Immunolocalization and visualization of the vWF pattern by the sequential incubation of the blot with a) primary vWF antiserum, b) peroxidase- or beta-galactosidase-conjugated secondary antibodies and a relevant chromogenic substrate.

Electrochemiluminescence Biosensor for Detection of Thrombin Using Enzyme-based Signal Amplification

2013 ◽  
Vol 40 (10) ◽  
pp. 1549-1554
Author(s):  
Hua-Yu ZHU ◽  
Li ZHANG ◽  
Huai-Cheng CHEN ◽  
Sheng-Juan YAN
Keyword(s):  
Signal Amplification
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