Accessing Molecular Dynamics in Cells by Fluorescence Correlation Spectroscopy
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Petra Dittrich
Abstract
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. Performed within diffractionlimited confocal volume elements, FCS provides an attractive alternative to photobleaching recovery methods for determining intracellular mobility parameters of very low quantities of fluorophores. Due to its high sensitivity sufficient for single molecule detection, the method is subject to certain artifact hazards that must be carefully controlled, such as photobleaching and intramolecular dynamics, which introduce fluorescence flickering. Furthermore, if molecular mobility is to be probed, nonspecific interactions of the labeling dye with cellular structures can introduce systematic errors. In cytosolic measurements, lipophilic dyes, such as certain rhodamines that bind to intracellular membranes, should be avoided. To study free diffusion, genetically encoded fluorescent labels such as green fluorescent protein (GFP) or DsRed are preferable since they are less likely to nonspecifically interact with cellular substructures.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Highlight: Fluorescence Correlation Spectroscopy
- Heuristic Statistical Analysis of Fluorescence Fluctuation Data with Bright Spikes: Application to Ligand Binding to the Human Serotonin Receptor Expressed in Escherichia coli Cells
- Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films
- Ligand-Receptor Interactions in the Membrane of Cultured Cells Monitored by Fluorescence Correlation Spectroscopy
- Fluorescence Fluctuation Analysis for the Study of Interactions between Oligonucleotides and Polycationic Polymers
- Determination of the Net Exchange Rate of Tubulin Dimer in Steady-State Microtubules by Fluorescence Correlation Spectroscopy
- UV-Fluorescence Correlation Spectroscopy of 2-Aminopurine
- Isolation of Two cDNAs Encoding Functional Human Cytoplasmic Cysteinyl-tRNA Synthetase
- Involvement of Intracellular Ca2+in the Regulation of Bovine Leukemia Virus Expression
- Nucleotide-Binding Sites in the Functional Unit of Sarcoplasmic Reticulum Ca2+-ATPase as Studied by Photoaffinity Spin-Labeled 2-N3-SL-ATP
- Interaction between Lipopolysaccharide (LPS), LPS-Binding Protein (LBP), and Planar Membranes
- Differential Binding of Urokinase and Peptide Antagonists to the Urokinase Receptor: Evidence from Characterization of the Receptor in Four Primate Species
- Effects of Cysteine to Serine Substitutions in the Two Inter-Chain Disulfide Bonds of Insulin
- Tricorn Protease in Bacteria: Characterization of the Enzyme from Streptomyces coelicolor
- Structures of Tryparedoxins Revealing Interaction with Trypanothione
- Quantitative Two-Color Fluorescence Cross-Correlation Spectroscopy in the Analysis of Polymerase Chain Reaction
- Fluorescence Correlation Spectroscopy as a New Method for the Investigation of Aptamer/Target Interactions
- Localization of Lysobisphosphatidic Acid-Rich Membrane Domains in Late Endosomes
- Fluorescence Correlation Spectroscopy for the Characterisation of Drug Delivery Systems
- Accessing Molecular Dynamics in Cells by Fluorescence Correlation Spectroscopy
- Tailor-Made Dyes for Fluorescence Correlation Spectroscopy (FCS)
- Synergistic Inhibition of the Glucocorticoid Receptor by Radicicol and Benzoquinone Ansamycins
Articles in the same Issue
- Highlight: Fluorescence Correlation Spectroscopy
- Heuristic Statistical Analysis of Fluorescence Fluctuation Data with Bright Spikes: Application to Ligand Binding to the Human Serotonin Receptor Expressed in Escherichia coli Cells
- Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films
- Ligand-Receptor Interactions in the Membrane of Cultured Cells Monitored by Fluorescence Correlation Spectroscopy
- Fluorescence Fluctuation Analysis for the Study of Interactions between Oligonucleotides and Polycationic Polymers
- Determination of the Net Exchange Rate of Tubulin Dimer in Steady-State Microtubules by Fluorescence Correlation Spectroscopy
- UV-Fluorescence Correlation Spectroscopy of 2-Aminopurine
- Isolation of Two cDNAs Encoding Functional Human Cytoplasmic Cysteinyl-tRNA Synthetase
- Involvement of Intracellular Ca2+in the Regulation of Bovine Leukemia Virus Expression
- Nucleotide-Binding Sites in the Functional Unit of Sarcoplasmic Reticulum Ca2+-ATPase as Studied by Photoaffinity Spin-Labeled 2-N3-SL-ATP
- Interaction between Lipopolysaccharide (LPS), LPS-Binding Protein (LBP), and Planar Membranes
- Differential Binding of Urokinase and Peptide Antagonists to the Urokinase Receptor: Evidence from Characterization of the Receptor in Four Primate Species
- Effects of Cysteine to Serine Substitutions in the Two Inter-Chain Disulfide Bonds of Insulin
- Tricorn Protease in Bacteria: Characterization of the Enzyme from Streptomyces coelicolor
- Structures of Tryparedoxins Revealing Interaction with Trypanothione
- Quantitative Two-Color Fluorescence Cross-Correlation Spectroscopy in the Analysis of Polymerase Chain Reaction
- Fluorescence Correlation Spectroscopy as a New Method for the Investigation of Aptamer/Target Interactions
- Localization of Lysobisphosphatidic Acid-Rich Membrane Domains in Late Endosomes
- Fluorescence Correlation Spectroscopy for the Characterisation of Drug Delivery Systems
- Accessing Molecular Dynamics in Cells by Fluorescence Correlation Spectroscopy
- Tailor-Made Dyes for Fluorescence Correlation Spectroscopy (FCS)
- Synergistic Inhibition of the Glucocorticoid Receptor by Radicicol and Benzoquinone Ansamycins
