scholarly journals Elimination of autofluorescence in fluorescence correlation spectroscopy using the AzaDiOxaTriAngulenium (ADOTA) fluorophore in combination with time-correlated single-photon counting (TCSPC)

2013 ◽  
Vol 405 (14) ◽  
pp. 4887-4894 ◽  
Author(s):  
Ryan M. Rich ◽  
Mark Mummert ◽  
Zygmunt Gryczynski ◽  
Julian Borejdo ◽  
Thomas Just Sørensen ◽  
...  
2019 ◽  
Vol 10 (1) ◽  
Author(s):  
James W. P. Brown ◽  
Arnaud Bauer ◽  
Mark E Polinkovsky ◽  
Akshay Bhumkar ◽  
Dominic J. B. Hunter ◽  
...  

AbstractSingle-molecule assays have, by definition, the ultimate sensitivity and represent the next frontier in biological analysis and diagnostics. However, many of these powerful technologies require dedicated laboratories and trained personnel and have therefore remained research tools for specialists. Here, we present a single-molecule confocal system built from a 3D-printed scaffold, resulting in a compact, plug and play device called the AttoBright. This device performs single photon counting and fluorescence correlation spectroscopy (FCS) in a simple format and is widely applicable to the detection of single fluorophores, proteins, liposomes or bacteria. The power of single-molecule detection is demonstrated by detecting single α-synuclein amyloid fibrils, that are currently evaluated as biomarkers for Parkinson’s disease, with an improved sensitivity of >100,000-fold over bulk measurements.


2015 ◽  
Vol 32 (6) ◽  
pp. 958-960 ◽  
Author(s):  
Dominic Waithe ◽  
Mathias P. Clausen ◽  
Erdinc Sezgin ◽  
Christian Eggeling

Abstract Motivation: Fluorescence Correlation Spectroscopy (FCS) is a popular tool for measuring molecular mobility and how mobility relates to molecular interaction dynamics and bioactivity in living cells. The FCS technique has been significantly advanced by its combination with super-resolution STED microscopy (STED-FCS). Specifically, the use of gated detection has shown great potential for enhancing STED-FCS, but has also created a demand for software which is efficient and also implements the latest algorithms. Prior to this study, no open software has been available which would allow practical time-gating and correlation of point data derived from STED-FCS experiments. Results: The product of this study is a piece of stand-alone software called FoCuS-point. FoCuS-point utilizes advanced time-correlated single-photon counting (TCSPC) correlation algorithms along with time-gated filtering and innovative data visualization. The software has been designed to be highly user-friendly and is tailored to handle batches of data with tools designed to process files in bulk. FoCuS-point also includes advanced fitting algorithms which allow the parameters of the correlation curves and thus the kinetics of diffusion to be established quickly and efficiently. Availability and implementation: FoCuS-point is written in python and is available through the github repository: https://github.com/dwaithe/FCS_point_correlator. Furthermore, compiled versions of the code are available as executables which can be run directly in Linux, Windows and Mac OSX operating systems. Contact: [email protected]


2014 ◽  
Vol 22 (23) ◽  
pp. 28783 ◽  
Author(s):  
Taro Yamashita ◽  
Dengkuan Liu ◽  
Shigehito Miki ◽  
Johtaro Yamamoto ◽  
Tokuko Haraguchi ◽  
...  

2003 ◽  
Vol 11 (26) ◽  
pp. 3583 ◽  
Author(s):  
Michael Wahl ◽  
Ingo Gregor ◽  
Mattias Patting ◽  
Jörg Enderlein

2019 ◽  
Author(s):  
Ipsita Saha ◽  
Saveez Saffarian

AbstractWe present a method utilizing single photon interference and fluorescence correlation spectroscopy (FCS) to simultaneously measure transport of fluorescent molecules within aqueous samples. Our method, within seconds, measures transport in thousands of homogenous voxels (100 nm)3 and eliminates photo-physical artifacts associated with blinking of fluorescent molecules. A comprehensive theoretical framework is presented and validated by measuring transport of quantum dots, associated with VSV-G receptor along cellular membranes as well as within viscous gels.


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