A Comprehensive Review of Fluorescence Correlation Spectroscopy

Ying Ma,Lan Yu,Juanjuan Zheng,Yunze Lei,Dan Dan,Peng Gao,Min Liu

doi:10.3389/fphy.2021.644450

Abstract

Fluorescence correlation spectroscopy (FCS) is a powerful technique for quantification of molecular dynamics, and it has been widely applied in diverse fields, e.g., biomedicine, biophysics, and chemistry. By time-correlation of the fluorescence fluctuations induced by molecules diffusing through a focused light, FCS can quantitatively evaluate the concentration, diffusion coefficient, and interaction of the molecules in vitro or in vivo. In this review, the basic principle and implementation of FCS are introduced. Then, the advances of FCS variants are reviewed, covering dual-color FCCS, multi-focus FCS, pair correlation function (pCF), scanning FCS, focus-reduced FCS, SPIM-FCS, and inverse-FCS. Besides, the applications of FCS are demonstrated with the measurement of local concentration, hydrodynamic radius, diffusion coefficient, and the interaction of different molecules. Lastly, a discussion is given by summarizing the pros and cons of different FCS techniques, as well as the outlooks and perspectives of FCS.

Highlights

A Comprehensive Review of Fluorescence Correlation SpectroscopyLan Yu 1†, Yunze Lei 1†, Ying Ma 1, Min Liu 1, Juanjuan Zheng 1, Dan Dan 2 and Peng Gao 1*
Molecular dynamics in biological systems are the foundation of life events
Inverse-Fluorescence correlation spectroscopy (FCS) measures unlabeled particles in a dense fluorescent medium, provides particle size and concentration independent of the diffusional dynamic. This method is only applicable to relatively large particles

Summary

A Comprehensive Review of Fluorescence Correlation Spectroscopy

Lan Yu 1†, Yunze Lei 1†, Ying Ma 1, Min Liu 1, Juanjuan Zheng 1, Dan Dan 2 and Peng Gao 1*. Fluorescence correlation spectroscopy (FCS) is a powerful technique for quantification of molecular dynamics, and it has been widely applied in diverse fields, e.g., biomedicine, biophysics, and chemistry. By time-correlation of the fluorescence fluctuations induced by molecules diffusing through a focused light, FCS can quantitatively evaluate the concentration, diffusion coefficient, and interaction of the molecules in vitro or in vivo. The basic principle and implementation of FCS are introduced. The advances of FCS variants are reviewed, covering dual-color FCCS, multi-focus FCS, pair correlation function (pCF), scanning FCS, focus-reduced FCS, SPIM-FCS, and inverseFCS. The applications of FCS are demonstrated with the measurement of local concentration, hydrodynamic radius, diffusion coefficient, and the interaction of different molecules. A discussion is given by summarizing the pros and cons of different FCS techniques, as well as the outlooks and perspectives of FCS

INTRODUCTION

Experimental Setup

SUMMARY