Fluorescence correlation spectroscopy for the detection and study of single molecules in biology.

  title={Fluorescence correlation spectroscopy for the detection and study of single molecules in biology.},
  author={Miguel {\'A}ngel Medina and Petra Schwille},
  journal={BioEssays : news and reviews in molecular, cellular and developmental biology},
  volume={24 8},
  • M. Medina, P. Schwille
  • Published 1 August 2002
  • Chemistry, Physics, Biology
  • BioEssays : news and reviews in molecular, cellular and developmental biology
The recent development of single molecule detection techniques has opened new horizons for the study of individual macromolecules under physiological conditions. Conformational subpopulations, internal dynamics and activity of single biomolecules, parameters that have so far been hidden in large ensemble averages, are now being unveiled. Herein, we review a particular attractive solution-based single molecule technique, fluorescence correlation spectroscopy (FCS). This time-averaging… 

Figures and Tables from this paper

Fluorescence correlation spectroscopy is a non-invasive technique and is based on fluctuations of fluorescence within a tiny observation volume under equilibrium conditions. With its wide
Single-Molecule Tracking and Its Application in Biomolecular Binding Detection
Various SMT techniques developed to date are discussed, with an emphasis on the recent development of the next generation 3D tracking system that not only achieves ultrahigh spatiotemporal resolution but also provides sufficient working depth suitable for live animal imaging.
Ultrafast Single-Molecule Spectroscopy
The detection and spectroscopic studies of individual molecules has begun to find widespread applications in the life sciences. While conventional time-resolved experiments continue to provide useful
Breaking the Concentration Barrier for Single-Molecule Fluorescence Measurements.
This Concept focuses on reviewing fundamental principles of single-molecule fluorescence techniques and wish to inspire development of new and better tools to achieve this goal.
Fluorescence correlation spectroscopy of autofluorescent proteins and its applications in live cell membranes
Fluorescence correlation spectroscopy (FCS) is used for investigating the kinetic processes and molecular interactions by means of the statistical analysis of fluorescence fluctuations. These
Single-Molecule Studies of Environmental Ionic-Strength Sensors using Fluorescence Correlation Spectroscopy (FCS)
Fluorescence Correlation Spectroscopy (FCS) is a power, single-molecule technique used in applied (high throughput screening of drugs) and basic (protein diffusion, structural conformation,
Diffusion Theory for Cell Membrane Fluorescence Microscopy
  • Minchul Kang
  • Biology
    Fluorescence Methods for Investigation of Living Cells and Microorganisms
  • 2020
This chapter will review some topics of the diffusion theory from both deterministic and probabilistic approaches, which are relevant to cell membrane fluorescence microscopy, and derive some basic equations for FARP and FCS based on the diffusion Theory.
Microfluidic Analysis and Parallel Confocal Detection of Single Molecules
The detection of single molecules using confocal spectroscopy is of major interest and tremendous improvements have been made within the last decade. The underlying basics, however, a pinhole and a


Fluorescence spectroscopy of single biomolecules.
The progress in applying single-molecule detection and single-Molecule spectroscopy at room temperature by laser-induced fluorescence with the use of fluorophores that are site-specifically attached to macromolecules is reviewed.
Accessing Molecular Dynamics in Cells by Fluorescence Correlation Spectroscopy
FCS analyzes spontaneous fluctuations in the fluorescence emission of small molecular ensembles, thus providing information about a multitude of parameters, such as concentrations, molecular mobility and dynamics of fluorescently labeled molecules, which provides an attractive alternative for determining intracellular mobility parameters of very low quantities of fluorophores.
Dual-color confocal fluorescence spectroscopy and its application in biotechnology. Fluorescence correlation Spectroscopy.
In recent years, fluorescence correlation spectroscopy (FCS) has become an attractive analytical tool for the investigation of biomolecular processes at the single molecular level. This method was
Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis.
It is demonstrated that two-photon excitation combined with dual-color fluorescence correlation spectroscopy can be the key to simplifying simultaneous investigations of multiple fluorescent species significantly on a single-molecule scale.
Fluorescence correlation spectroscopy : theory and applications
FCS in environmental analysis.- New outlooks for FCS.- FCS in nucleic acid analysis.- FCS in high throughput screening.- Signal analysis in FCS.- FCS in the study of protein-ligand interaction.- FCS
Rapid characterization of green fluorescent protein fusion proteins on the molecular and cellular level by fluorescence correlation microscopy.
Fluorescence correlation microscopy was applied to characterize fusion proteins of the green fluorescent protein (GFP) on the cellular as well as molecular level within seconds in an integrated instrument, offering new possibilities for highly integrated high-throughput screening.
Sorting single molecules: application to diagnostics and evolutionary biotechnology.
  • M. Eigen, R. Rigler
  • Biology, Physics
    Proceedings of the National Academy of Sciences of the United States of America
  • 1994
The method expands the horizon in molecular diagnostics by making it possible to monitor concentrations down to (less than) 10(-15) M without any need for amplification.