Verfügbarkeit: Optical imaging techniques in cell biology

Optical imaging techniques in cell biology:

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Format:	Buch
Sprache:	English
Veröffentlicht:	Boca Raton, Fla. [u.a.] CRC Press 2012
Ausgabe:	2., rev. ed.
Schlagworte:	Mikroskopie Cytologie
Online-Zugang:	Inhaltsverzeichnis Klappentext
Beschreibung:	Literaturangaben
Beschreibung:	XVII, 297 S. zahlr. Ill. und graph. Darst.
ISBN:	9781439848258 1439848254

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MARC


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Datensatz im Suchindex

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adam_text	Contents Introduction: The Optical Microscope in Cell Biology ............................................xi Contributors .............................................................................................................xv Acknowledgments ..................................................................................................xvii Chapter 1 The Light Microscope ..........................................................................1 Lenses and Microscopes ......................................................................1 The Back Focal Plane of a Lens ......................................................4 Good Resolution ..............................................................................8 Resolution: Rayleigh s Approach ................................................9 Abbe ..........................................................................................13 Add a Drop of Oil .........................................................................16 Köhler Illumination .......................................................................18 References ..........................................................................................19 Chapter 2 Optical Contrasting Techniques .........................................................21 Darkfield .............................................................................................21 Phase Contrast ....................................................................................23 Polarization ........................................................................................25 Differential Interference Contrast ......................................................28 Hoffman Modulation Contrast ...........................................................31 Which Technique Is Best? ..................................................................34 References ..........................................................................................34 Chapter 3 Fluorescence and Fluorescence Microscopy ......................................35 What Is Fluorescence? ........................................................................35 What Makes a Molecule Fluorescent? ..........................................39 The Fluorescence Microscope ...........................................................40 Optical Arrangement .....................................................................40 Light Source ..............................................................................40 Filter Sets: Excitation Filter, Dichroic Mirror, and Barrier Filter ..........................................................................................43 References ..........................................................................................47 Chapter 4 Image Capture ....................................................................................49 Optical Layout for Image Capture .....................................................49 Color Recording .................................................................................49 Additive Color Model ....................................................................50 Subtractive Color Model ................................................................51 VI Contents CCD Cameras.....................................................................................53 Frame-Transfer Array ....................................................................55 Interline-Transfer Array ................................................................55 Back Illumination ..........................................................................56 Binning ..........................................................................................56 Capturing Color ..................................................................................57 Filter Wheels .................................................................................57 Filter Mosaics ................................................................................58 Three CCD Elements with Dichroic Beamsplitters ......................58 Boosting the Signal ............................................................................58 References ..........................................................................................61 Chapter 5 The Confocal Microscope ..................................................................63 The Scanning Optical Microscope .....................................................63 The Confocal Principle ......................................................................63 Resolution and Point Spread Function ...............................................65 Lateral Resolution in the Confocal Microscope ............................68 Practical Confocal Microscopes ........................................................69 The Light Source: Lasers ..............................................................71 Gas Lasers .................................................................................72 Solid-State Lasers .....................................................................72 Semiconductor Lasers ...............................................................73 Supercontinuum Lasers ............................................................74 Laser Delivery ...............................................................................75 The Primary Beamsplitter ........................................................76 Beam Scanning .........................................................................77 Pinhole and Signal Channel Configurations .............................77 Detectors ........................................................................................81 References ..........................................................................................82 Further Reading .................................................................................82 Chapter 6 The Digital Image ..............................................................................83 Pixels and Voxels ................................................................................83 Contrast ..............................................................................................83 Spatial Sampling: TheNyquist Criterion ...........................................85 Temporal Sampling: Signal-to-Noise Ratio .......................................86 Multichannel Images ..........................................................................89 References ..........................................................................................91 Further Reading .................................................................................91 Chapter 7 Aberrations and Their Consequences ................................................93 Geometrical Aberrations ....................................................................93 Spherical Aberration .....................................................................93 Contents Coma ..............................................................................................99 Astigmatism .................................................................................100 Field Curvature ............................................................................100 Chromatic Aberration ......................................................................102 Chromatic Difference of Magnification ......................................103 Practical Consequences ....................................................................105 Apparent Depth ...........................................................................107 References ........................................................................................108 Further Reading ...............................................................................108 Chapter 8 Nonlinear Microscopy ......................................................................109 Multiphoton Microscopy ..................................................................109 Principles of Two-Photon Fluorescence ......................................109 Theory and Practice ................................................................ Ill Lasers for Nonlinear Microscopy ...........................................112 Advantages of Two-Photon Excitation ...................................114 Construction of a Multiphoton Microscope .....................................115 Fluorochromes for Multiphoton Microscopy ..............................116 Second Harmonic Microscopy .........................................................117 Summary ..........................................................................................122 References ........................................................................................122 Further Reading ...............................................................................123 Chapter 9 High-Speed Confocal Microscopy ...................................................125 Tandem Scanning (Spinning Disk) Microscopes .............................125 Petrán System ..............................................................................125 One-Sided Tandem Scanning Microscopes (OTSMS) ................128 Microlens Array: The Yokogawa System ....................................128 Slit-Scanning Microscopes ...............................................................130 Multipoint-Array Scanners ...............................................................132 Structured Illumination ....................................................................135 References ........................................................................................135 Further Reading ...............................................................................136 Chapter 10 Deconvolution and Image Processing ..............................................137 Deconvolution ..................................................................................137 Deconvolving Confocal Images ..................................................142 Image Processing .............................................................................142 Grayscale Operations ..................................................................143 Image Arithmetic ........................................................................145 Convolution: Smoothing and Sharpening ...................................146 References ........................................................................................149 Further Reading ...............................................................................149 viii Contents Chapter 11 Three-Dimensional Imaging: Stereoscopy and Reconstruction ......151 Surfaces: Two-and-a-Half Dimensions ............................................151 Perception of the 3D World ..............................................................153 Motion Parallax ...........................................................................153 Convergence and Focus of Our Eyes ...........................................153 Perspective ...................................................................................154 Concealment of One Object by Another .....................................154 Our Knowledge of the Size and Shape of Everyday Things .......154 Light and Shade ...........................................................................154 Limitations of Confocal Microscopy ...............................................154 Stereoscopy ......................................................................................155 Three-Dimensional Reconstruction .................................................157 Techniques That Require Identification of Objects .................158 Techniques That Create Views Directly from Intensity Data .....161 Simple Projections ..................................................................161 Weighted Projection (Alpha Blending) ...................................163 References ........................................................................................168 Chapter 12 Green Fluorescent Protein ................................................................169 Structure and Properties of GFP ......................................................169 GFP Variants ....................................................................................170 Applications of GFP .........................................................................175 Heat Shock ...................................................................................176 Cationic Lipid Reagents ..............................................................176 DEAE-Dextran and Polybrene ...................................................176 Calcium Phosphate Coprecipitation .............................................176 Electroporation ............................................................................177 Microinjection .............................................................................177 Gene Gun .....................................................................................177 Plants: Agrobacterium .................................................................179 References ........................................................................................180 Chapter 13 Fluorescent Staining .........................................................................181 Teresa Dibbayawan, Eleanor Kable, and Guy Cox Immunolabeling ...............................................................................181 Types of Antibody ............................................................................182 Raising Antibodies ...........................................................................183 Labeling ............................................................................................185 Fluorescent Stains for Cell Components and Compartments ..........187 References ........................................................................................190 Chapter 14 Quantitative Fluorescence ................................................................191 Fluorescence Intensity Measurements .............................................191 Linearity Calibration ...................................................................191 Contents ix Measurement ...............................................................................192 Colocalization..............................................................................192 Ratio Imaging.............................................................................. 195 Cell Loading ................................................................................199 Membrane Potential .........................................................................199 Fast-Response Dyes .....................................................................200 Slow-Response Dyes ....................................................................200 Fluorescence Recovery after Photobleaching .............................201 References ........................................................................................205 Chapter 15 Advanced Fluorescence Techniques: FLIM, FRET, and FCS .........207 Fluorescence Lifetime ......................................................................207 Practical Lifetime Microscopy (FLIM) ...........................................209 Frequency Domain ......................................................................209 Time Domain ...............................................................................209 Fluorescence Resonant Energy Transfer (FRET) ............................213 Why Use FRET? .........................................................................214 Identifying and Quantifying FRET .............................................215 Increase in Brightness of Acceptor Emission .........................215 Quenching of Emission from the Donor .................................219 Lifetime of Donor Emission ...................................................220 Protection from Bleaching of Donor ......................................221 Fluorescence Correlation Spectroscopy (FCS) ................................223 Raster Image Correlation Spectroscopy ...........................................224 References ........................................................................................225 Further Reading ...............................................................................226 Chapter 16 Evanescent Wave Microscopy ..........................................................227 The Near-Field and Evanescent Waves ............................................227 Total Internal Reflection Microscopy ..........................................227 Near-Field Microscopy .....................................................................230 References ........................................................................................236 Chapter 17 Beyond the Diffraction Limit ...........................................................237 4Pi and Multiple-Objective Microscopy ..........................................237 Stimulated Emission Depletion (STED) ...........................................240 Structured Illumination ....................................................................245 Stochastic Techniques ......................................................................247 Super-Resolution Summary .............................................................252 References ........................................................................................253 Appendix A: Microscope Care and Maintenance ............................................255 Cleaning ...........................................................................................256 The Fluorescent Illuminator .............................................................259 x Contents Appendix В: Keeping Cells Alive under the Microscope .................................261 Eleanor Kable and Guy Cox Chambers ..........................................................................................261 Light .................................................................................................265 Movement .........................................................................................265 Finally ...............................................................................................265 Appendix C: Antibody Labeling of Plant and Animal Cells: Tips and Sample Schedules .................................................................................................267 Eleanor Kable and Teresa Dibbayawan Antibodies: Tips on Handling and Storage ......................................267 Pipettes: Tips on Handling ..........................................................268 Antibodies and Antibody Titrations ............................................268 Example ..................................................................................268 Immunofluorescence Protocol ..........................................................270 Method .........................................................................................270 Multiple Labeling and Different Samples ...................................271 Plant Material ..............................................................................271 Protocol ...................................................................................272 Diagram Showing Position of Antibodies on Multiwell Slide... 273 Appendix D: Image Processing with ImageJ ....................................................275 Nuno Moreno Introduction ......................................................................................275 Different Windows in ImageJ ..........................................................275 Image Levels ................................................................................276 Colors and Look-Up Tables .........................................................276 Size Calibration ................................................................................277 Image Math ......................................................................................279 Quantification ...................................................................................280 Stacks and 3D Representation ..........................................................280 FFT and Image Processing ..............................................................282 Macro Language in ImageJ ..............................................................283 Index ......................................................................................................................285 íl îmag irig ТесЛ л Zgt/eş;/^^ immunofluorescence and fluorescent proteins — which have revolutionized cell biology. §»АШ*Ш%задкаж»ршїШКА ■ИИ tttt№ ■UU MU Яві mm IHM are accurate and detailed, but as far as possible non-mathematical. This edition includes appendices with useful practical protocols, references, and suggestions for further reading.
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spelling	Optical imaging techniques in cell biology Guy Cox 2., rev. ed. Boca Raton, Fla. [u.a.] CRC Press 2012 XVII, 297 S. zahlr. Ill. und graph. Darst. txt rdacontent n rdamedia nc rdacarrier Literaturangaben Mikroskopie (DE-588)4039238-7 gnd rswk-swf Cytologie (DE-588)4070177-3 gnd rswk-swf Cytologie (DE-588)4070177-3 s Mikroskopie (DE-588)4039238-7 s DE-604 Cox, Guy Christopher 1945- Sonstige (DE-588)133149056 oth Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024779185&sequence=000003&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024779185&sequence=000004&line_number=0002&func_code=DB_RECORDS&service_type=MEDIA Klappentext
spellingShingle	Optical imaging techniques in cell biology Mikroskopie (DE-588)4039238-7 gnd Cytologie (DE-588)4070177-3 gnd
subject_GND	(DE-588)4039238-7 (DE-588)4070177-3
title	Optical imaging techniques in cell biology
title_auth	Optical imaging techniques in cell biology
title_exact_search	Optical imaging techniques in cell biology
title_full	Optical imaging techniques in cell biology Guy Cox
title_fullStr	Optical imaging techniques in cell biology Guy Cox
title_full_unstemmed	Optical imaging techniques in cell biology Guy Cox
title_short	Optical imaging techniques in cell biology
title_sort	optical imaging techniques in cell biology
topic	Mikroskopie (DE-588)4039238-7 gnd Cytologie (DE-588)4070177-3 gnd
topic_facet	Mikroskopie Cytologie
url	http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024779185&sequence=000003&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024779185&sequence=000004&line_number=0002&func_code=DB_RECORDS&service_type=MEDIA
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