Comparison between time-correlated single-photon counting and fluorescence correlation spectroscopy in single-molecule identification

1998 ◽  
Author(s):  
Joerg Enderlein ◽  
Malte Koellner
Keyword(s):  
Single Molecule ◽  
Fluorescence Correlation Spectroscopy ◽  
Single Photon ◽  
Photon Counting ◽  
Correlation Spectroscopy ◽  
Single Photon Counting ◽  
Fluorescence Correlation

scholarly journals Single-molecule detection on a portable 3D-printed microscope

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
James W. P. Brown ◽  
Arnaud Bauer ◽  
Mark E Polinkovsky ◽  
Akshay Bhumkar ◽  
Dominic J. B. Hunter ◽  
...  
Keyword(s):  
Single Molecule ◽  
Amyloid Fibrils ◽  
Single Photon ◽  
Photon Counting ◽  
Correlation Spectroscopy ◽  
Single Molecule Detection ◽  
Single Photon Counting ◽  
Fluorescence Correlation ◽  
3D Printed ◽  
Simple Format

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.

scholarly journals FoCuS-point: software for STED fluorescence correlation and time-gated single photon counting

2015 ◽  
Vol 32 (6) ◽  
pp. 958-960 ◽  
Author(s):  
Dominic Waithe ◽  
Mathias P. Clausen ◽  
Erdinc Sezgin ◽  
Christian Eggeling
Keyword(s):  
Single Photon ◽  
Photon Counting ◽  
Super Resolution ◽  
Correlation Spectroscopy ◽  
Single Photon Counting ◽  
Fluorescence Correlation ◽  
Point Data ◽  
User Friendly ◽  
Kinetics Of ◽  
Focus Point

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]

Fluorescence correlation spectroscopy: The technique and its applications in soft matter

2018 ◽  
Vol 4 (4) ◽  
Author(s):  
Anjali Gupta ◽  
Jagadish Sankaran ◽  
Thorsten Wohland
Keyword(s):  
Single Molecule ◽  
Fluorescence Correlation Spectroscopy ◽  
Soft Matter ◽  
Photophysical Properties ◽  
Spatiotemporal Analysis ◽  
Correlation Spectroscopy ◽  
Basic Principles ◽  
Chemical Reaction Kinetics ◽  
Fluorescence Correlation ◽  
Wide Range

Abstract Fluorescence correlation spectroscopy (FCS) is a well-established single-molecule method used for the quantitative spatiotemporal analysis of dynamic processes in a wide range of samples. It possesses single-molecule sensitivity but provides ensemble averaged molecular parameters such as mobility, concentration, chemical reaction kinetics, photophysical properties and interaction properties. These parameters have been utilized to characterize a variety of soft matter systems. This review provides an overview of the basic principles of various FCS modalities, their instrumentation, data analysis, and the applications of FCS to soft matter systems.

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