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Quantitative Single-Molecule Electrochemiluminescence Bioassay |
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| Authors: | Wenxin Zhu Jinrun Dong Guoxiang Ruan Yuan Zhou Prof Jiandong Feng |
| Institution: | 1. Laboratory of Experimental Physical Biology, Department of Chemistry, Zhejiang University, Hangzhou, 310027 China
These authors contributed equally to this work. Contribution: Data curation (lead), Formal analysis (lead), Investigation (lead), Methodology (lead), Validation (equal), Visualization (lead), Writing - original draft (equal), Writing - review & editing (supporting);2. Laboratory of Experimental Physical Biology, Department of Chemistry, Zhejiang University, Hangzhou, 310027 China These authors contributed equally to this work. Contribution: Data curation (equal), Formal analysis (equal), Investigation (equal), Methodology (equal), Validation (equal);3. Department of Laboratory Medicine, Key Laboratory of Clinical In Vitro Diagnostic Techniques of Zhejiang Province, Institute of Laboratory Medicine, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310003 China Contribution: Data curation (supporting), Formal analysis (supporting), Resources (lead), Validation (supporting);4. Laboratory of Experimental Physical Biology, Department of Chemistry, Zhejiang University, Hangzhou, 310027 China Contribution: Investigation (supporting), Validation (lead);5. Laboratory of Experimental Physical Biology, Department of Chemistry, Zhejiang University, Hangzhou, 310027 China |
| Abstract: | A single-molecule electrochemiluminescence bioassay is developed here which allows imaging and direct quantification of single biomolecules. Imaging single biomolecules is realized by localizing the electrochemiluminescence events of the labeled molecules. Such an imaging system allows mapping the spatial distribution of biomolecules with electrochemiluminescence and contains quantitative single-molecule insights. We further quantify biomolecules by spatiotemporally merging the repeated reactions at one molecule site and then counting the clustered molecules. The proposed single-molecule electrochemiluminescence bioassay is used to detect carcinoembryonic antigen, showing a limit of detection of 67 attomole concentration which is 10 000 times better than conventional electrochemiluminescence bioassays. This spatial resolution and sensitivity enable single-molecule electrochemiluminescence bioassay a new toolbox for both specific bioimaging and ultrasensitive quantitative analysis. |
| Keywords: | Electrochemiluminescence Quantitative Imaging Single-Molecule Bioassay Super-Resolution Microscopy |