scholarly journals High throughput single molecule detection for monitoring biochemical reactions

The Analyst ◽  
2009 ◽  
Vol 134 (1) ◽  
pp. 97-106 ◽  
Author(s):  
Paul I. Okagbare ◽  
Steven A. Soper
Author(s):  
Xiaojia Jiang ◽  
Mingsong Zang ◽  
Fei Li ◽  
Chunxi Hou ◽  
Quan Luo ◽  
...  

Biological nanopore-based techniques have attracted more and more attention recently in the field of single-molecule detection, because they allow the real-time, sensitive, high-throughput analysis. Herein, we report an engineered biological...


2003 ◽  
Vol 8 (1) ◽  
pp. 19-33 ◽  
Author(s):  
Ulrich Haupts ◽  
Martin Rüdiger ◽  
Stephen Ashman ◽  
Sandra Turconi ◽  
Ryan Bingham ◽  
...  

Single-molecule detection technologies are becoming a powerful readout format to support ultra-high-throughput screening. These methods are based on the analysis of fluorescence intensity fluctuations detected from a small confocal volume element. The fluctuating signal contains information about the mass and brightness of the different species in a mixture. The authors demonstrate a number of applications of fluorescence intensity distribution analysis (FIDA), which discriminates molecules by their specific brightness. Examples for assays based on brightness changes induced by quenching/dequenching of fluorescence, fluorescence energy transfer, and multiple-binding stoichiometry are given for important drug targets such as kinases and proteases. FIDA also provides a powerful method to extract correct biological data in the presence of compound fluorescence. ( Journal of Biomolecular Screening 2003:19-33)


Lab on a Chip ◽  
2019 ◽  
Vol 19 (1) ◽  
pp. 79-86 ◽  
Author(s):  
Mattia Fontana ◽  
Carel Fijen ◽  
Serge G. Lemay ◽  
Klaus Mathwig ◽  
Johannes Hohlbein

Single-molecule detection schemes offer powerful means to overcome static and dynamic heterogeneity inherent to complex samples.


1999 ◽  
Vol 4 (6) ◽  
pp. 335-353 ◽  
Author(s):  
Keith J. Moore ◽  
Sandra Turconi ◽  
Stephen Ashman ◽  
Martin Ruediger ◽  
Ulrich Haupts ◽  
...  

Fluorescence assay technologies used for miniaturized high throughput screening are broadly divided into two classes. Macroscopic fluorescence techniques (encompassing conventional fluorescence intensity, anisotropy [also often referred to as fluorescence polarization] and energy transfer) monitor the assay volume- and time-averaged fluorescence output from the ensemble of emitting fluorophores. In contrast, single-molecule detection (SMD) techniques and related approaches, such as fluorescence correlation spectroscopy (FCS), stochastically sample the fluorescence properties of individual constituent molecules and only then average many such detection events to define the properties of the assay system as a whole. Analysis of single molecular events is accomplished using confocal optics with an illumination/detection volume of -1 fl (10-15 L) such that the signal is insensitive to miniaturization of HTS assays to 1 A1 or below. In this report we demonstrate the general applicability of one SMD technique (FCS) to assay configuration for target classes typically encountered in HTS and confirm the equivalence of the rate/equilibrium constants determined by FCS and by macroscopic techniques. Advantages and limitations of the current FCS technology, as applied here, and potential solutions, particularly involving alternative SMD detection techniques, are also discussed.


2021 ◽  
Author(s):  
Yang Liu ◽  
Ye Deng ◽  
Yanmei Yang ◽  
Yuanyuan Qu ◽  
Chao Zhang ◽  
...  

Solid-state nanopore detection and sequencing of single-molecule offers a new paradigm because of several well-recognized features like long-read, high throughput, high precision and direct analyses. However, several key technical challenges...


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