UID:
almafu_9958130093802883
Umfang:
1 online resource (l, 325 pages, 18 unnumbered pages of plates) :
,
illustrations (some color).
ISBN:
9781283936446
,
1283936445
,
9780124058576
,
0124058574
Serie:
Methods in enzymology ; volume five hundred and nineteen
Inhalt:
This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers fluorescence fluctuation spectroscopy and includes chapters on such topics as Förster resonance energy transfer (fret) with fluctuation algorithms, protein corona on nanoparticles by FCS, and FFS approaches to the study of receptors in live cells. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers fluorescence fluctuation spectrosco
Anmerkung:
"ISSN: 0076-6879."
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Front Cover; Fluorescence Fluctuation Spectroscopy (FFS), Part B; Copyright; Contents; Contributors; Preface; Methods in Enzymology; Chapter One: FCS in STED Microscopy: Studying the Nanoscale of Lipid Membrane Dynamics; 1. Introduction; 2. STED-FCS of Lipid Membrane Dynamics; 2.1. Cellular labeling; 2.2. STED nanoscopy; 2.3. STED-FCS: The principle; 2.4. STED-FCS: Anomalous subdiffusion; 2.5. STED-FCS: Quantification of anomalous subdiffusion; 3. Lipid Membrane Dynamics; 3.1. Anomaly of PE diffusion; 3.2. Consistency of the different analysis procedures; 3.3. Environmental parameters
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3.4. Molecular dependence3.5. Dependence on COase treatment; 4. Conclusions; Acknowledgments; References; Chapter Two: Analyzing Förster Resonance Energy Transfer with Fluctuation Algorithms; 1. Introduction; 1.1. Definition of the correlation functions; 1.2. Heterogeneous mixtures; 1.3. Hardware; 1.4. Timescales studied by FRET; 2. FRET and FCS; 2.1. Only FRET molecules; 2.2. FRET molecules mixed with donor-only sample; 3. Filtered FCS; 3.1. FRET-fFCS: FRET molecules mixed with donor-only at picomolar concentration; 3.2. FRET and fFCS interconversion between states at SMD; 4. Applications
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5. Discussion5.1. Relaxation time; 5.1.1. Temporal boundaries for FRET-FCS; 5.1.2. Temporal boundaries for fFCS; 5.1.3. Alternative and complementary methods; 5.2. Brightnesses uncertainty; 5.3. Advantages of fFCS; Acknowledgments; References; Chapter Three: Fluorescence Fluctuation Spectroscopy Approaches to the Study of Receptors in Live Cells; 1. Introduction; 2. Selected FFS Studies; 2.1. Determination of receptor densities; 2.2. Measurement of binding affinities; 2.3. Receptor oligomerization state and clustering; 2.4. Analysis of nuclear receptors
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3. Choice of Fluorophores: General Considerations3.1. Fluorescent antibodies; 3.2. Fluorescent proteins; 3.3. Biomolecular fluorescence complementation; 3.4. HaloTags/SNAP/FlAsH; 3.5. Quantum dots; 4. Cells: General Considerations; 4.1. Cell growth and transfection; 4.2. Maintenance of cell viability; 4.3. Photobleaching and phototoxicity; 4.4. Autofluorescence; 5. Summary; Acknowledgments; References; Chapter Four: Studying the Protein Corona on Nanoparticles by FCS; 1. Introduction; 2. Sample Preparation; 2.1. Proteins; 2.2. Nanoparticles
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2.3. Preparation of a protein concentration series by serial dilution2.4. Sample cell and sample loading; 3. Experimental Procedures; 3.1. Experimental setups; 3.2. Microscope calibration; 3.3. Data collection; 4. Data Analysis; 4.1. Correlation function analysis; 4.2. Computation of hydrodynamic radii; 5. Protein Corona Formation Measured by FCS; 5.1. Concentration dependence of protein adhesion; 5.2. Structure of the protein corona; 5.3. Protein electrostatics and adsorption tendency; 6. Conclusions; Acknowledgments; References
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Chapter Five: Studying Antibody-Antigen Interactions with Fluorescence Fluctuation Spectroscopy
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English
Weitere Ausg.:
ISBN 9780124055391
Weitere Ausg.:
ISBN 0124055397
Sprache:
Englisch
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