ArticleClinical chemistry2012
Four-color alternating-laser excitation single-molecule fluorescence spectroscopy for next-generation biodetection assays.
Article in Clinical chemistry, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
17 citing papers in PubMed, 33 citations in OpenAlex.
- ATP-driven conformational dynamics reveal hidden intermediates in a heterodimeric ABC transporter.eLife · 2026Article
- Parallel stopped-flow interrogation of diverse biological systems at the single-molecule scale.Nature methods · 2026Article
- DNA-FRET Constructs Enable Multiplexed Fluorescence Detection at the Single-Molecule Level.Chemical & biomedical imaging · 2024Article
- Single-molecule fluorescence multiplexing by multi-parameter spectroscopic detection of nanostructured FRET labels.Nature nanotechnology · 2024Article
- Multiplexed smFRET Nucleic Acid Sensing Using DNA Nanotweezers.Biosensors · 2023Article
- Multi-parameter photon-by-photon hidden Markov modeling.Nature communications · 2022Article
- FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices.eLife · 2021Review
- Advances in Optical Single-Molecule Detection: En Route to Supersensitive Bioaffinity Assays.Angewandte Chemie (International ed. in English) · 2020Review
- High-throughput multiplexed autoantibody detection to screen type 1 diabetes and multiple autoimmune diseases simultaneously.EBioMedicine · 2019Article
- Multiplex Single-Molecule DNA Barcoding Using an Oligonucleotide Ligation Assay.Biophysical journal · 2018Article
- 48-spot single-molecule FRET setup with periodic acceptor excitation.The Journal of chemical physics · 2018Article
- Toward dynamic structural biology: Two decades of single-molecule Förster resonance energy transfer.Science (New York, N.Y.) · 2018Review
- Förster resonance energy transfer and protein-induced fluorescence enhancement as synergetic multi-scale molecular rulers.Scientific reports · 2016Article
- A Quantitative Theoretical Framework For Protein-Induced Fluorescence Enhancement-Förster-Type Resonance Energy Transfer (PIFE-FRET).The journal of physical chemistry. B · 2016Article
- Silicon photon-counting avalanche diodes for single-molecule fluorescence spectroscopy.IEEE journal of selected topics in quantum electronics : a publication of the IEEE Lasers and Electro-optics Society · 2014Article
- Non-Coding RNAs: Functional Aspects and Diagnostic Utility in Oncology.International journal of molecular sciences · 2013Article
- Development of new photon-counting detectors for single-molecule fluorescence microscopy.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2013Review
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
Abstract
backgroundSingle-molecule detection (SMD) technologies are well suited for clinical diagnostic applications by offering the prospect of minimizing precious patient sample requirements while maximizing clinical information content. Not yet available, however, is a universal SMD-based platform technology that permits multiplexed detection of both nucleic acid and protein targets and that is suitable for automation and integration into the clinical laboratory work flow.
methodsWe have used a sensitive, specific, quantitative, and cost-effective homogeneous SMD method that has high single-well multiplexing potential and uses alternating-laser excitation (ALEX) fluorescence-aided molecule sorting extended to 4 colors (4c-ALEX). Recognition molecules are tagged with different-color fluorescence dyes, and coincident confocal detection of ≥2 colors constitutes a positive target-detection event. The virtual exclusion of the majority of sources of background noise eliminates washing steps. Sorting molecules with multidimensional probe stoichiometries (S) and single-molecule fluorescence resonance energy transfer efficiencies (E) allows differentiation of numerous targets simultaneously.
resultsWe show detection, differentiation, and quantification-in a single well-of (a) 25 different fluorescently labeled DNAs; (b) 8 bacterial genetic markers, including 3 antibiotic drug-resistance determinants found in 11 septicemia-causing Staphylococcus and Enterococcus strains; and (c) 6 tumor markers present in blood.
conclusionsThe results demonstrate assay utility for clinical molecular diagnostic applications by means of multiplexed detection of nucleic acids and proteins and suggest potential uses for early diagnosis of cancer and infectious and other diseases, as well as for personalized medicine. Future integration of additional technology components to minimize preanalytical sample manipulation while maximizing throughput should allow development of a user-friendly ("sample in, answer out") point-of-care platform for next-generation medical diagnostic tests that offer considerable savings in costs and patient sample.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.