Biophysical methods in cell biology / edited by Ewa K. Paluch.

Contributor(s): Paluch, Ewa K [editor.]
Material type: TextTextSeries: Methods in cell biology: v. 125.Publisher: Amsterdam : Elsevier Ltd., [2015]Copyright date: ©2015Edition: First editionDescription: 1 online resource (xxvi, 512 pages) : illustrations (some color)Content type: text Media type: computer Carrier type: online resourceISBN: 9780128013267; 0128013265; 1322871442; 9781322871448Subject(s): Biophysics | Cell nuclei | DNA | RNA | SCIENCE / Life Sciences / Anatomy & Physiology | Biophysics | Cell nuclei | DNA | RNA | Cell Biology -- methods -- Laboratory Manuals | Biophysics -- methods -- Laboratory Manuals | Cell Biology | Cytological Techniques | Biophysics -- methodsGenre/Form: Electronic books. | Electronic books. DDC classification: 571.4 LOC classification: QH505Online resources: Click here to access online
Contents:
Front Cover; Methods in Cell Biology; Series Editors; Methods in Cell Biology ; Copyright ; Contents; Contributors; Preface; 1. Single-molecule imaging of cytoplasmic dynein in vivo; Introduction; 1. Visualization of Cytoplasmic Dynein In Vivo; 1.1 Background; 1.2 Experiment; 1.2.1 Preparation of fission yeast zygotes; 1.2.2 Observation of dyneins in the cytoplasm; 1.2.3 Observation of dyneins on the microtubule; 1.2.4 Discussion; 1.2.4.1 Prebleaching to observe dyneins with higher SNR; 1.2.4.1 Prebleaching to observe dyneins with higher SNR; 1.2.4.2 Estimation of penetration depth of HILO.
1.2.4.2 Estimation of penetration depth of HILO2. Image Analysis; 2.1 Tracking of Single Molecules; 2.2 Confirmation of Single-Molecule Imaging; 2.3 Analysis of Dynein Movement; 3. Conclusion; 4. Methods; 4.1 Cell Culture; 4.2 Preparation of Samples for Imaging; 4.3 Microscopy; References; 2. Single-molecule imaging in live cell using gold nanoparticles; Introduction and Rationale; 1. Gold Nanoparticle Synthesis and Functionalization; 1.1 Materials; 1.1.1 List of chemicals required for the nanoparticle synthesis and functionalization; 1.2 Nanoparticle Synthesis.
1.3 Nanoparticle Functionalization with Nanobodies1.4 Sample Characterization; 1.4.1 Absorption spectra; 1.4.2 Transmission electron microscope; 1.4.3 Agarose gel electrophoresis; 2. Photothermal Imaging; 2.1 Materials; 2.2 Principle; 2.3 Experimental Setup; 2.4 Resolution and Sensitivity; 2.4.1 Resolution; 2.4.2 Sensitivity; 3. Live Cell Imaging; 3.1 Cell surface labeling; 3.2 2D Single-Particle Tracking; 3.3 Particle Internalization; Conclusion; Acknowledgments; References; 3. Quantitative measurement of transcription dynamics in living cells; 1. Visualizing Transcription in Living Cells.
2. Experimental Protocols3. Cell Segmentation; 4. Measuring Spot Intensity; 5. Correcting for Background MS2-GFP Level; 6. Tracking Algorithms; 7. Additional Cell Properties; 8. Summary; References; 4. An easy-to-use single-molecule speckle microscopy enabling nanometer-scale flow and wide-range lifetime measurement of cell ...; Introduction; 1. Methods; 1.1 Preparation of Actin Probes; 1.1.1 Required materials; 1.1.2 Procedure for DyLight NHS ester labeling of actin; 1.2 Electroporation Method for Delivery of DL-Actin to XTC Cells; 1.2.1 Required materials.
1.2.2 Procedure for electroporation of DL-actin into XTC cells1.3 SiMS Imaging; 1.3.1 Required materials; 1.3.2 Examples of microscopy setups; 1.3.3 Procedure for the SiMS imaging of DL-actin loaded XTC cells; 1.4 Data Analysis; 1.4.1 Nanometer-scale displacement measurement; 1.4.2 Simultaneous analysis of actin dynamics with diverse timescales; 2. Perspectives; Acknowledgments; References; 5. Dissecting microtubule structures by laser ablation; Introduction; 1. Theoretical Framework; 2. Microtubule Organization Measurements; 2.1 Extract and Sample Preparation.
Summary: "This new volume of Methods in Cell Biology looks at methods for analyzing of biophysical methods in cell biology. Chapters cover such topics as AFM, traction force microscopy, digital holographic microscopy, single molecule imaging, video force microscopy and 3D multicolor super-resolution screening."-- Provided by publisher.
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Includes bibliographical references and index.

Description based on online resource; title from PDF title page (ScienceDirect, viewed February 11, 2015).

"This new volume of Methods in Cell Biology looks at methods for analyzing of biophysical methods in cell biology. Chapters cover such topics as AFM, traction force microscopy, digital holographic microscopy, single molecule imaging, video force microscopy and 3D multicolor super-resolution screening."-- Provided by publisher.

Front Cover; Methods in Cell Biology; Series Editors; Methods in Cell Biology ; Copyright ; Contents; Contributors; Preface; 1. Single-molecule imaging of cytoplasmic dynein in vivo; Introduction; 1. Visualization of Cytoplasmic Dynein In Vivo; 1.1 Background; 1.2 Experiment; 1.2.1 Preparation of fission yeast zygotes; 1.2.2 Observation of dyneins in the cytoplasm; 1.2.3 Observation of dyneins on the microtubule; 1.2.4 Discussion; 1.2.4.1 Prebleaching to observe dyneins with higher SNR; 1.2.4.1 Prebleaching to observe dyneins with higher SNR; 1.2.4.2 Estimation of penetration depth of HILO.

1.2.4.2 Estimation of penetration depth of HILO2. Image Analysis; 2.1 Tracking of Single Molecules; 2.2 Confirmation of Single-Molecule Imaging; 2.3 Analysis of Dynein Movement; 3. Conclusion; 4. Methods; 4.1 Cell Culture; 4.2 Preparation of Samples for Imaging; 4.3 Microscopy; References; 2. Single-molecule imaging in live cell using gold nanoparticles; Introduction and Rationale; 1. Gold Nanoparticle Synthesis and Functionalization; 1.1 Materials; 1.1.1 List of chemicals required for the nanoparticle synthesis and functionalization; 1.2 Nanoparticle Synthesis.

1.3 Nanoparticle Functionalization with Nanobodies1.4 Sample Characterization; 1.4.1 Absorption spectra; 1.4.2 Transmission electron microscope; 1.4.3 Agarose gel electrophoresis; 2. Photothermal Imaging; 2.1 Materials; 2.2 Principle; 2.3 Experimental Setup; 2.4 Resolution and Sensitivity; 2.4.1 Resolution; 2.4.2 Sensitivity; 3. Live Cell Imaging; 3.1 Cell surface labeling; 3.2 2D Single-Particle Tracking; 3.3 Particle Internalization; Conclusion; Acknowledgments; References; 3. Quantitative measurement of transcription dynamics in living cells; 1. Visualizing Transcription in Living Cells.

2. Experimental Protocols3. Cell Segmentation; 4. Measuring Spot Intensity; 5. Correcting for Background MS2-GFP Level; 6. Tracking Algorithms; 7. Additional Cell Properties; 8. Summary; References; 4. An easy-to-use single-molecule speckle microscopy enabling nanometer-scale flow and wide-range lifetime measurement of cell ...; Introduction; 1. Methods; 1.1 Preparation of Actin Probes; 1.1.1 Required materials; 1.1.2 Procedure for DyLight NHS ester labeling of actin; 1.2 Electroporation Method for Delivery of DL-Actin to XTC Cells; 1.2.1 Required materials.

1.2.2 Procedure for electroporation of DL-actin into XTC cells1.3 SiMS Imaging; 1.3.1 Required materials; 1.3.2 Examples of microscopy setups; 1.3.3 Procedure for the SiMS imaging of DL-actin loaded XTC cells; 1.4 Data Analysis; 1.4.1 Nanometer-scale displacement measurement; 1.4.2 Simultaneous analysis of actin dynamics with diverse timescales; 2. Perspectives; Acknowledgments; References; 5. Dissecting microtubule structures by laser ablation; Introduction; 1. Theoretical Framework; 2. Microtubule Organization Measurements; 2.1 Extract and Sample Preparation.

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