Incorporation of STED technique into single-molecule spectroscopy to break the concentration limit of diffusing molecules in single-molecule detection

2018 ◽  
Vol 54 (69) ◽  
pp. 9667-9670
Author(s):  
Namdoo Kim ◽  
Jiwoong Kwon ◽  
Youngbin Lim ◽  
Jooyoun Kang ◽  
Sohyeon Bae ◽  
...  
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection ◽  
Concentration Limit ◽  
Single Molecule Spectroscopy

Incorporation of STED into ALEX-FRET increases the concentration limit of single-molecule detection by 100-fold to 5 nM.

scholarly journals Single molecule fingerprinting reveals different amplification properties of α-synuclein oligomers and preformed fibrils in seeding assay.

2021 ◽  
Author(s):  
Derrick Lau ◽  
Chloe Magnan ◽  
Kathryn Hill ◽  
Antony Cooper ◽  
Yann Gambin ◽  
...  
Keyword(s):  
Single Molecule ◽  
Amyloid Fibrils ◽  
Size Exclusion ◽  
Single Molecule Detection ◽  
Single Molecule Spectroscopy ◽  
Parkinsons Disease ◽  
Exclusion Chromatography ◽  
Particle Level ◽  
Amplification Assay ◽  
Multiple Species

The quantification of α-synuclein (α-syn) aggregates has emerged as a promising biomarker for synucleinopathies. Assays that amplify and detect such aggregates have revealed the presence of seeding-competent species in biosamples of patients diagnosed with Parkinsons disease. However, multiple species such as oligomers and amyloid fibrils, are formed during the aggregation of α-synuclein and these species are likely to co-exist in biological samples and thus it remains unclear which species(s) are contributing to the signal detected in seeding assays. To identify which species can be detected in seeding assays, recombinant oligomers and preformed fibrils were produced and purified to characterise their individual biochemical and seeding potential. Here, we used single molecule spectroscopy to track the formation and purification of oligomers and fibrils at the single particle level and compare their respective seeding potential in an amplification assay. Single molecule detection validates that size-exclusion chromatography efficiently separates oligomers from fibrils. Oligomers were found to be seeding-competent but our results reveal that their seeding behaviour is very different compared to preformed fibrils in our amplification assay. Overall, our data suggest that even a low number of preformed fibrils present in biosamples are likely to dominate the response in seeding assays.

Breaking the concentration limit of optical single-molecule detection

2014 ◽  
Vol 43 (4) ◽  
pp. 1014-1028 ◽  
Author(s):  
Phil Holzmeister ◽  
Guillermo P. Acuna ◽  
Dina Grohmann ◽  
Philip Tinnefeld
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection ◽  
Concentration Limit

scholarly journals Recent advances in plasmonic nanocavities for single-molecule spectroscopy

2021 ◽  
Author(s):  
Nicolò Maccaferri ◽  
Grégory Barbillon ◽  
Alemayehu Nana Koya ◽  
Guowei Lu ◽  
Guillermo P. Acuna ◽  
...  
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection ◽  
Single Molecule Spectroscopy ◽  
Recent Advances ◽  
Review Reports

This mini-review reports the most recent results on plasmonic nanocavities applied to enhanced single-molecule detection and spectroscopy.

Use of single-molecule spectroscopy to tackle fundamental problems in biochemistry: using studies on purple bacterial antenna complexes as an example

2009 ◽  
Vol 422 (2) ◽  
pp. 193-205 ◽  
Author(s):  
Richard J. Cogdell ◽  
Jürgen Köhler
Keyword(s):  
Electronic Structure ◽  
Energy Transfer ◽  
Single Molecule ◽  
Physical Structure ◽  
Single Molecule Detection ◽  
Transfer Reactions ◽  
Single Molecule Spectroscopy

Optical single-molecule techniques can be used in two modes to investigate fundamental questions in biochemistry, namely single-molecule detection and single-molecule spectroscopy. This review provides an overview of how single-molecule spectroscopy can be used to gain detailed information on the electronic structure of purple bacterial antenna complexes and to draw conclusions about the underlying physical structure. This information can be used to understand the energy-transfer reactions that are responsible for the earliest reactions in photosynthesis.

ChemInform Abstract: Breaking the Concentration Limit of Optical Single-Molecule Detection

ChemInform ◽  
2014 ◽  
Vol 45 (15) ◽  
pp. no-no
Author(s):  
Phil Holzmeister ◽  
Guillermo P. Acuna ◽  
Dina Grohmann ◽  
Philip Tinnefeld
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection ◽  
Concentration Limit

scholarly journals A controlled T7 transcription-driven symmetric amplification cascade machinery for single-molecule detection of multiple repair glycosylases

2021 ◽  
Author(s):  
Li-juan Wang ◽  
Le Liang ◽  
Bing-jie Liu ◽  
BingHua Jiang ◽  
Chun-yang Zhang
Keyword(s):  
Single Molecule ◽  
Single Molecule Detection ◽  
Amplification Cascade

A controlled T7 transcription-driven symmetric amplification cascade machinery is developed for single-molecule detection of multiple repair glycosylases.

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