Petra Schwille

Zwei-Photonen-Kreuzkorrelations-Spektroskopie : Nachweis der Interaktionen einzelner Moleküle in der lebenden Zelle (9999)

Prof. Dr. Rer. Nat. Petra Schwille

The progress of miniaturisation towards the nanoscopic scale in science and technology has also influenced the biosciences. This is particularly important, since proteins, as the smallest functional...

Supercritical angle fluorescence correlation spectroscopy (2007)

Ries, Jonas, Ruckstuhl, Thomas, Verdes, Dorinel, Schwille, Petra

We explore the potential of a supercritical angle (SA) objective for fluorescence correlation spectroscopy (FCS). This novel microscope objective combines tight focusing by an aspheric lens with...

Probing Lipid Mobility of Raft-exhibiting Model Membranes by Fluorescence Correlation Spectroscopy (2003)

Poolman, Bert, Bacia, Kirsten, Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

Confocal fluorescence microscopy and fluorescence correlation spectroscopy (FCS) have been employed to investigate the lipid spatial and dynamic organization in giant unilamellar vesicles (GUVs)...

Characterization of lipid bilayer phases by confocal microscopy and fluorescence correlation spectroscopy

Korlach, Jonas, Schwille, Petra, Webb, Watt W., Feigenson, Gerald W.

We report the application of confocal imaging and fluorescence correlation spectroscopy (FCS) to characterize chemically well-defined lipid bilayer models for biomembranes. Giant unilamellar vesicles...

Real-time enzyme kinetics monitored by dual-color fluorescence cross-correlation spectroscopy

Kettling, Ulrich, Koltermann, Andre, Schwille, Petra, Eigen, Manfred

A method for sensitively monitoring enzyme kinetics and activities by using dual-color fluorescence cross-correlation spectroscopy is described. This universal method enables the development of...

Fluorescence correlation analysis of probe diffusion simplifies quantitative pathogen detection by PCR

Walter, Nils G., Schwille, Petra, Eigen, Manfred

A sensitive, labor-saving, and easily automatable nonradioactive procedure named APEX-FCS (amplified probe extension detected by fluorescence correlation spectroscopy) has been established to detect...

Detection of HIV-1 RNA by nucleic acid sequence-based amplification combined with fluorescence correlation spectroscopy

Oehlenschläger, Frank, Schwille, Petra, Eigen, Manfred

Nucleic acid sequence-based amplification (NASBA) has proved to be an ultrasensitive method for HIV-1 diagnosis in plasma even in the primary HIV infection stage. This technique was combined with...

Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy

Haupts, Ulrich, Maiti, Sudipta, Schwille, Petra, Webb, Watt W.

We have investigated the pH dependence of the dynamics of conformational fluctuations of green fluorescent protein mutants EGFP (F64L/S65T) and GFP-S65T in small ensembles of molecules in solution by...

Fluorescence correlation spectroscopy reveals fast optical excitation-driven intramolecular dynamics of yellow fluorescent proteins

Schwille, Petra, Kummer, Susanne, Heikal, Ahmed A., Moerner, W. E., Webb, Watt W.

Fast excitation-driven fluctuations in the fluorescence emission of yellow-shifted green fluorescent protein mutants T203Y and T203F, with S65G/S72A, are discovered in the 10−6–10−3-s time...

Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis

Heinze, Katrin G., Koltermann, Andre, Schwille, Petra

Confocal fluorescence correlation spectroscopy as a time-averaging fluctuation analysis combining maximum sensitivity with high statistical confidence has proved to be a very versatile and powerful...

A protease assay for two-photon crosscorrelation and FRET analysis based solely on fluorescent proteins

Kohl, Tobias, Heinze, Katrin G., Kuhlemann, Rene, Koltermann, Andre, Schwille, Petra

GFP and the red fluorescent protein, DsRed, have been combined to design a protease assay that allows not only for fluorescence resonance energy transfer (FRET) studies but also for dual-color...

Intracellular calmodulin availability accessed with two-photon cross-correlation

Kim, Sally A., Heinze, Katrin G., Waxham, M. Neal, Schwille, Petra

The availability and interactions of signaling proteins are tightly regulated in time and space to produce specific and localized effects. For calmodulin (CaM), a key transducer of intracellular Ca2+...

Sterol structure determines the separation of phases and the curvature of the liquid-ordered phase in model membranes

Bacia, Kirsten, Schwille, Petra, Kurzchalia, Teymuras

The existence of lipid rafts in biological membranes in vivo is still debated. In contrast, the formation of domains in model systems has been well documented. In giant unilamellar vesicles (GUVs)...

An ultrasensitive site-specific DNA recombination assay based on dual-color fluorescence cross-correlation spectroscopy

Jahnz, Michael, Schwille, Petra

Site-specific exchange of genetic information is mediated by DNA recombinases, such as FLP or Cre, and has become a valuable tool in modern molecular biology. The so far low number of suitable...

Probing the endocytic pathway in live cells using dual-color fluorescence cross-correlation analysis.

Bacia, Kirsten, Majoul, Irina V, Schwille, Petra

Fluorescence (auto)correlation spectroscopy (FCS) has developed into a widely used method for investigating molecular dynamics and mobility of molecules in vitro and in vivo. Dual-color...

Two-photon fluorescence coincidence analysis: rapid measurements of enzyme kinetics.

Heinze, Katrin G, Rarbach, Markus, Jahnz, Michael, Schwille, Petra

Dual-color fluorescence cross-correlation analysis is a powerful tool for probing interactions of different fluorescently labeled molecules in aqueous solution. The concept is the selective...

Lipid Dynamics and Domain Formation in Model Membranes Composed of Ternary Mixtures of Unsaturated and Saturated Phosphatidylcholines and Cholesterol

Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

In recent years, the implication of sphingomyelin in lipid raft formation has intensified the long sustained interest in this membrane lipid. Accumulating evidences show that cholesterol...

Triple-Color Coincidence Analysis: One Step Further in Following Higher Order Molecular Complex Formation

Heinze, Katrin G., Jahnz, Michael, Schwille, Petra

Confocal fluorescence spectroscopy is a versatile method for studying dynamics and interactions of biomolecules in their native environment with minimal interference with the observed system....

Fluorescence Correlation Spectroscopy Relates Rafts in Model and Native Membranes

Bacia, Kirsten, Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

The lipid raft model has evoked a new perspective on membrane biology. Understanding the structure and dynamics of lipid domains could be a key to many crucial membrane-associated processes in cells....

The Mobility of Phytochrome within Protonemal Tip Cells of the Moss Ceratodon purpureus, Monitored by Fluorescence Correlation Spectroscopy

Böse, Guido, Schwille, Petra, Lamparter, Tilman

Fluorescence correlation spectroscopy (FCS) is a versatile tool for investigating the mobilities of fluorescent molecules in cells. In this article, we show that it is possible to distinguish between...

Characterization of Interaction between Cationic Lipid-Oligonucleotide Complexes and Cellular Membrane Lipids Using Confocal Imaging and Fluorescence Correlation Spectroscopy

Gordon, Sean Patrick, Berezhna, Svitlana, Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

Complexes formed by cationic liposomes and single-strand oligodeoxynucleotides (CL-ODN) are promising delivery systems for antisense therapy. ODN release from the complexes is an essential step for...

In situ fluorescence analysis demonstrates active siRNA exclusion from the nucleus by Exportin 5

Ohrt, Thomas, Merkle, Dennis, Birkenfeld, Karin, Echeverri, Christophe J., Schwille, Petra

Two types of short double-stranded RNA molecules, namely microRNAs (miRNAs) and short interfering RNAs (siRNAs), have emerged recently as important regulators of gene expression. Although these...

Two-Photon Cross-Correlation Analysis of Intracellular Reactions with Variable Stoichiometry

Kim, Sally A., Heinze, Katrin G., Bacia, Kirsten, Waxham, M. Neal, Schwille, Petra

We successfully demonstrate the effectiveness of two-photon fluorescence cross-correlation spectroscopy (TPCCS) to study the complex binding stoichiometry of calmodulin (CaM) and Ca2+/CaM-dependent...

Determining Protease Activity In Vivo by Fluorescence Cross-Correlation Analysis

Kohl, Tobias, Haustein, Elke, Schwille, Petra

To date, most biochemical approaches to unravel protein function have focused on purified proteins in vitro. Whereas they analyze enzyme performance under assay conditions, they do not necessarily...

Characterization of lipid bilayer phases by confocal microscopy and fluorescence correlation spectroscopy

Korlach, Jonas, Schwille, Petra, Webb, Watt W., Feigenson, Gerald W.

We report the application of confocal imaging and fluorescence correlation spectroscopy (FCS) to characterize chemically well-defined lipid bilayer models for biomembranes. Giant unilamellar vesicles...

Real-time enzyme kinetics monitored by dual-color fluorescence cross-correlation spectroscopy

Kettling, Ulrich, Koltermann, Andre, Schwille, Petra, Eigen, Manfred

A method for sensitively monitoring enzyme kinetics and activities by using dual-color fluorescence cross-correlation spectroscopy is described. This universal method enables the development of...

Fluorescence correlation analysis of probe diffusion simplifies quantitative pathogen detection by PCR

Walter, Nils G., Schwille, Petra, Eigen, Manfred

A sensitive, labor-saving, and easily automatable nonradioactive procedure named APEX-FCS (amplified probe extension detected by fluorescence correlation spectroscopy) has been established to detect...

Detection of HIV-1 RNA by nucleic acid sequence-based amplification combined with fluorescence correlation spectroscopy

Oehlenschläger, Frank, Schwille, Petra, Eigen, Manfred

Nucleic acid sequence-based amplification (NASBA) has proved to be an ultrasensitive method for HIV-1 diagnosis in plasma even in the primary HIV infection stage. This technique was combined with...

Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy

Haupts, Ulrich, Maiti, Sudipta, Schwille, Petra, Webb, Watt W.

We have investigated the pH dependence of the dynamics of conformational fluctuations of green fluorescent protein mutants EGFP (F64L/S65T) and GFP-S65T in small ensembles of molecules in solution by...

Fluorescence correlation spectroscopy reveals fast optical excitation-driven intramolecular dynamics of yellow fluorescent proteins

Schwille, Petra, Kummer, Susanne, Heikal, Ahmed A., Moerner, W. E., Webb, Watt W.

Fast excitation-driven fluctuations in the fluorescence emission of yellow-shifted green fluorescent protein mutants T203Y and T203F, with S65G/S72A, are discovered in the 10−6–10−3-s time...

Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis

Heinze, Katrin G., Koltermann, Andre, Schwille, Petra

Confocal fluorescence correlation spectroscopy as a time-averaging fluctuation analysis combining maximum sensitivity with high statistical confidence has proved to be a very versatile and powerful...

A protease assay for two-photon crosscorrelation and FRET analysis based solely on fluorescent proteins

Kohl, Tobias, Heinze, Katrin G., Kuhlemann, Rene, Koltermann, Andre, Schwille, Petra

GFP and the red fluorescent protein, DsRed, have been combined to design a protease assay that allows not only for fluorescence resonance energy transfer (FRET) studies but also for dual-color...

Intracellular calmodulin availability accessed with two-photon cross-correlation

Kim, Sally A., Heinze, Katrin G., Waxham, M. Neal, Schwille, Petra

The availability and interactions of signaling proteins are tightly regulated in time and space to produce specific and localized effects. For calmodulin (CaM), a key transducer of intracellular Ca2+...

Sterol structure determines the separation of phases and the curvature of the liquid-ordered phase in model membranes

Bacia, Kirsten, Schwille, Petra, Kurzchalia, Teymuras

The existence of lipid rafts in biological membranes in vivo is still debated. In contrast, the formation of domains in model systems has been well documented. In giant unilamellar vesicles (GUVs)...

An ultrasensitive site-specific DNA recombination assay based on dual-color fluorescence cross-correlation spectroscopy

Jahnz, Michael, Schwille, Petra

Site-specific exchange of genetic information is mediated by DNA recombinases, such as FLP or Cre, and has become a valuable tool in modern molecular biology. The so far low number of suitable...

Probing the endocytic pathway in live cells using dual-color fluorescence cross-correlation analysis.

Bacia, Kirsten, Majoul, Irina V, Schwille, Petra

Fluorescence (auto)correlation spectroscopy (FCS) has developed into a widely used method for investigating molecular dynamics and mobility of molecules in vitro and in vivo. Dual-color...

Two-photon fluorescence coincidence analysis: rapid measurements of enzyme kinetics.

Heinze, Katrin G, Rarbach, Markus, Jahnz, Michael, Schwille, Petra

Dual-color fluorescence cross-correlation analysis is a powerful tool for probing interactions of different fluorescently labeled molecules in aqueous solution. The concept is the selective...

Lipid Dynamics and Domain Formation in Model Membranes Composed of Ternary Mixtures of Unsaturated and Saturated Phosphatidylcholines and Cholesterol

Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

In recent years, the implication of sphingomyelin in lipid raft formation has intensified the long sustained interest in this membrane lipid. Accumulating evidences show that cholesterol...

Triple-Color Coincidence Analysis: One Step Further in Following Higher Order Molecular Complex Formation

Heinze, Katrin G., Jahnz, Michael, Schwille, Petra

Confocal fluorescence spectroscopy is a versatile method for studying dynamics and interactions of biomolecules in their native environment with minimal interference with the observed system....

Fluorescence Correlation Spectroscopy Relates Rafts in Model and Native Membranes

Bacia, Kirsten, Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

The lipid raft model has evoked a new perspective on membrane biology. Understanding the structure and dynamics of lipid domains could be a key to many crucial membrane-associated processes in cells....

The Mobility of Phytochrome within Protonemal Tip Cells of the Moss Ceratodon purpureus, Monitored by Fluorescence Correlation Spectroscopy

Böse, Guido, Schwille, Petra, Lamparter, Tilman

Fluorescence correlation spectroscopy (FCS) is a versatile tool for investigating the mobilities of fluorescent molecules in cells. In this article, we show that it is possible to distinguish between...

Characterization of Interaction between Cationic Lipid-Oligonucleotide Complexes and Cellular Membrane Lipids Using Confocal Imaging and Fluorescence Correlation Spectroscopy

Gordon, Sean Patrick, Berezhna, Svitlana, Scherfeld, Dag, Kahya, Nicoletta, Schwille, Petra

Complexes formed by cationic liposomes and single-strand oligodeoxynucleotides (CL-ODN) are promising delivery systems for antisense therapy. ODN release from the complexes is an essential step for...

Two-Photon Cross-Correlation Analysis of Intracellular Reactions with Variable Stoichiometry

Kim, Sally A., Heinze, Katrin G., Bacia, Kirsten, Waxham, M. Neal, Schwille, Petra

We successfully demonstrate the effectiveness of two-photon fluorescence cross-correlation spectroscopy (TPCCS) to study the complex binding stoichiometry of calmodulin (CaM) and Ca2+/CaM-dependent...

Determining Protease Activity In Vivo by Fluorescence Cross-Correlation Analysis

Kohl, Tobias, Haustein, Elke, Schwille, Petra

To date, most biochemical approaches to unravel protein function have focused on purified proteins in vitro. Whereas they analyze enzyme performance under assay conditions, they do not necessarily...

In situ fluorescence analysis demonstrates active siRNA exclusion from the nucleus by Exportin 5

Ohrt, Thomas, Merkle, Dennis, Birkenfeld, Karin, Echeverri, Christophe J., Schwille, Petra

Two types of short double-stranded RNA molecules, namely microRNAs (miRNAs) and short interfering RNAs (siRNAs), have emerged recently as important regulators of gene expression. Although these...

Effects of Ceramide on Liquid-Ordered Domains Investigated by Simultaneous AFM and FCS

Chiantia, Salvatore, Kahya, Nicoletta, Ries, Jonas, Schwille, Petra

The sphingolipid ceramides are known to influence lipid lateral organization in biological membranes. In particular, ceramide-induced alterations of microdomains can be involved in several cell...

Studying Slow Membrane Dynamics with Continuous Wave Scanning Fluorescence Correlation Spectroscopy

Ries, Jonas, Schwille, Petra

Here we discuss the application of scanning fluorescence correlation spectroscopy (SFCS) using continuous wave excitation to analyze membrane dynamics. The high count rate per molecule enables the...

Phosphatidylethanolamine critically supports internalization of cell-penetrating protein C inhibitor

Baumgärtner, Petra, Geiger, Margarethe, Zieseniss, Susanne, Malleier, Julia, Huntington, James A., Hochrainer, Karin, ...

Although their contribution remains unclear, lipids may facilitate noncanonical routes of protein internalization into cells such as those used by cell-penetrating proteins. We show that protein C...

Pore Formation by a Bax-Derived Peptide: Effect on the Line Tension of the Membrane Probed by AFM

García-Sáez, Ana J., Chiantia, Salvatore, Salgado, Jesús, Schwille, Petra

Bax is a critical regulator of physiological cell death that increases the permeability of the outer mitochondrial membrane and facilitates the release of the so-called apoptotic factors during...

Fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy reveal the cytoplasmic origination of loaded nuclear RISC in vivo in human cells

Ohrt, Thomas, Mütze, Jörg, Staroske, Wolfgang, Weinmann, Lasse, Höck, Julia, Crell, Karin, ...

Studies of RNA interference (RNAi) provide evidence that in addition to the well-characterized cytoplasmic mechanisms, nuclear mechanisms also exist. The mechanism by which the nuclear RNA-induced...