Fluorescence microscopy: from principles to biological applications
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| Format: | Book |
| Language: | English |
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Weinheim
Wiley-VCH
[2017]
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| Edition: | Second edition |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | xxii, 482 Seiten Illustrationen, Diagramme (farbig) |
| ISBN: | 9783527338375 |
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| 245 | 1 | 0 | |a Fluorescence microscopy |b from principles to biological applications |c edited by Ulrich Kubitscheck |
| 250 | |a Second edition | ||
| 264 | 1 | |a Weinheim |b Wiley-VCH |c [2017] | |
| 264 | 4 | |c © 2017 | |
| 300 | |a xxii, 482 Seiten |b Illustrationen, Diagramme (farbig) | ||
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| _version_ | 1804176659938541568 |
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| adam_text | CONTENTS
LIST OF CONTRIBUTORS XV
PREFACE XIX
1 INTRODUCTION TO OPTICS 1
RAINER HEINTZMANN AND ULRICH KUBITSCHECK
1.1 A SHORT HISTORY OF THEORIES ABOUT LIGHT 1
1.2 PROPERTIES OF LIGHT WAVES 2
1.2.1 AN EXPERIMENT ON INTERFERENCE 2
1.2.2 PHYSICAL DESCRIPTION OF LIGHT WAVES 3
1.3 FOUR EFFECTS OF INTERFERENCE 7
1.3.1 DIFFRACTION 7
1.3.2 THE REFRACTIVE INDEX 9
1.3.3 REFRACTION 9
1.3.4 REFLECTION 10
1.3.5 LIGHT WAVES AND LIGHT RAYS 11
1.4 OPTICAL ELEMENTS 13
1.4.1 LENSES 13
1.4.2 METALLIC MIRRORS 15
1.4.3 DIELECTRIC MIRRORS 16
1.4.4 FILTERS 17
1.4.5 CHROMATIC REFLECTORS 18
1.5 OPTICAL ABERRATIONS 20
REFERENCES 22
2 PRINCIPLES OF LIGHT MICROSCOPY 23
ULRICH KUBITSCHECK
2.1 INTRODUCTION 23
2.2 CONSTRUCTION OF LIGHT MICROSCOPES 23
2.2.1 COMPONENTS OF LIGHT MICROSCOPES 23
2.2.2 IMAGING PATH 24
2.2.3 MAGNIFICATION 26
2.2.4 ANGULAR AND NUMERICAL APERTURE 27
2.2.5 FIELD OF VIEW 28
2.2.6 ILLUMINATION BEAM PATH 28
2.2.6.1 CRITICAL AND KOEHLER ILLUMINATION 28
2.2.6.2 BRIGHT-FIELD AND EPI-ILLUMINATION 31
2.3 WAVE OPTICS AND RESOLUTION 32
2.3.1 WAVE OPTICAL DESCRIPTION OF THE IMAGING PROCESS 33
2.3.2 THE AIRY PATTERN 37
2.3.3 POINT SPREAD FUNCTION AND OPTICAL TRANSFER FUNCTION 40
2.3.4 LATERAL AND AXIAL RESOLUTION 41
2.3.4.1 LATERAL RESOLUTION USING INCOHERENT LIGHT SOURCES 41
2.3.4.2 LATERAL RESOLUTION OF COHERENT LIGHT SOURCES 43
2.3A3 AXIAL RESOLUTION 45
2.3.5 MAGNIFICATION AND RESOLUTION 48
2.3.6 DEPTH OF FIELD AND DEPTH OF FOCUS 49
2.3.7 OVER- AND UNDERSAMPLING 50
2.4 APERTURES, PUPILS, AND TELECENTRICITY 50
2.5 MICROSCOPE OBJECTIVES 53
2.5.1 OBJECTIVE LENS DESIGN 53
2.5.2 LIGHT COLLECTION EFFICIENCY AND IMAGE BRIGHTNESS 57
2.5.3 OBJECTIVE LENS CLASSES 61
2.5.4 IMMERSION MEDIA 62
2.5.5 SPECIAL APPLICATIONS 65
2.6 CONTRAST 67
2.6.1 DARK FIELD 68
2.6.2 PHASE CONTRAST 69
2.6.2.1 FRITS ZERNIKE S EXPERIMENTS 70
2.6.2.2 SETUP OF A PHASE-CONTRAST MICROSCOPE 73
2.6.2.3 PROPERTIES OF PHASE-CONTRAST IMAGES 74
2.6.3 INTERFERENCE CONTRAST 74
2.6.4 ADVANCED TOPIC: DIFFERENTIAL INTERFERENCE CONTRAST 77
2.6.4.1 OPTICAL SETUP OF A DIC MICROSCOPE 77
2.6A2 INTERPRETATION OF DIC IMAGES 81
2.6A3 COMPARISON BETWEEN DIC AND PHASE CONTRAST 81
2.7 SUMMARY 82
ACKNOWLEDGMENTS 82
REFERENCES 83
3 FLUORESCENCE MICROSCOPY 85
JUREK W. DOBRUCKI AND ULRICH KUBITSCHECK
3.1 CONTRAST IN OPTICAL MICROSCOPY 85
3.2 PHYSICAL FOUNDATIONS OF FLUORESCENCE 86
3.2.1 WHAT IS FLUORESCENCE? 86
3.2.2 FLUORESCENCE EXCITATION AND EMISSION SPECTRA 89
3.3 FEATURES OF FLUORESCENCE MICROSCOPY 90
3.3.1 IMAGE CONTRAST 90
3.3.2 SPECIFICITY OF FLUORESCENCE LABELING 93
3.3.3 SENSITIVITY OF DETECTION 94
3.4 A FLUORESCENCE MICROSCOPE 95
3.4.1 PRINCIPLE OF OPERATION 95
3.4.2 SOURCES OF EXCITING LIGHT 99
3.4.3 OPTICAL FILTERS IN A FLUORESCENCE MICROSCOPE 101
3.4.4 ELECTRONIC FILTERS 103
3.4.5 PHOTODETECTORS FOR FLUORESCENCE MICROSCOPY 104
3.4.6 CCD OR CHARGE-COUPLED DEVICE 104
3.4.7 INTENSIFIED CCD (ICCD) 107
3.4.8 ELECTRON-MULTIPLYING CHARGE-COUPLED DEVICE (EMCCD) 109
3.4.9 CMOS 111
3.4.10 SCIENTIFIC CMOS (SCMOS) 112
3.4.11 FEATURES OF CCD AND CMOS CAMERAS 112
3.4.12 CHOOSING A DIGITAL CAMERA FOR FLUORESCENCE MICROSCOPY 113
3.4.13 PHOTOMULTIPLIER TUBE (PMT) 113
3.4.14 AVALANCHE PHOTODIODE (APD) 114
3.5 TYPES OF NOISE IN A DIGITAL MICROSCOPY IMAGE 114
3.6 QUANTITATIVE FLUORESCENCE MICROSCOPY 119
3.6.1 MEASUREMENTS OF FLUORESCENCE INTENSITY AND CONCENTRATION OF THE
LABELED TARGET 119
3.6.2 RATIOMETRIC MEASUREMENTS (CA++, PH) 121
3.6.3 MEASUREMENTS OF DIMENSIONS IN 3D FLUORESCENCE
MICROSCOPY 121
3.6.4 MEASUREMENTS OF EXCITING LIGHT INTENSITY 122
3.6.5 TECHNICAL TIPS FOR QUANTITATIVE FLUORESCENCE MICROSCOPY 123
3.7 LIMITATIONS OF FLUORESCENCE MICROSCOPY 124
3.7.1 PHOTOBLEACHING 124
3.7.2 REVERSIBLE PHOTOBLEACHING UNDER OXIDIZING OR REDUCING
CONDITIONS 125
3.7.3 PHOTOTOXICITY 125
3.7.4 OPTICAL RESOLUTION 126
3.7.5 MISREPRESENTATION OF SMALL OBJECTS 127
3.8 SUMMARY AND OUTLOOK 128
REFERENCES 130
RECOMMENDED INTERNET RESOURCES 131
FLUORESCENT SPECTRA DATABASE 132
4 FLUORESCENCE LABELING 133
GERD ULRICH NIENHAUS AND KARIN NIENHAUS
4.1 INTRODUCTION 133
4.2 KEY PROPERTIES OF FLUORESCENT LABELS 133
4.3 SYNTHETIC FLUOROPHORES 138
4.3.1 ORGANIC DYES 138
4.3.2 FLUORESCENT NANOPARTICLES 140
4.3.3 CONJUGATION STRATEGIES FOR SYNTHETIC FLUOROPHORES 142
4.3.4 NON-NATURAL AMINO ACIDS 146
4.3.5 BRINGING THE FLUOROPHORE TO ITS TARGET 147
4.4 GENETICALLY ENCODED LABELS 149
4.4.1 PHYCOBILIPROTEINS 149
4.4.2 GFP-LIKE PROTEINS 150
4.5 LABEL SELECTION FOR PARTICULAR APPLICATIONS 155
4.5.1 FRET TO MONITOR INTRAMOLECULAR CONFORMATIONAL DYNAMICS 155
4.5.2 PROTEIN EXPRESSION IN CELLS 159
4.5.3 FLUOROPHORES AS SENSORS INSIDE THE CELL 160
4.5.4 LIVE-CELL DYNAMICS 160
4.5.5 SUPER-RESOLUTION IMAGING 160
4.6 SUMMARY 161
REFERENCES 162
5 CONFOCAL MICROSCOPY 165
NIKOLAUS NAREDI-RAINER, JENS PRESCHER, ACHIM HARTSCHUH, AND
DON C.
LAMB
5.1 EVOLUTION AND LIMITS OF CONVENTIONAL WIDEFIELD MICROSCOPY 165
5.2 THEORY OF CONFOCAL MICROSCOPY 166
5.2.1 PRINCIPLE OF CONFOCAL MICROSCOPY 166
5.2.2 RADIAL AND AXIAL RESOLUTION AND THE IMPACT OF THE PINHOLE SIZE 173
5.2.3 SCANNING CONFOCAL IMAGING 179
5.2.3.1 STAGE SCANNING 179
5.2.3.2 LASER SCANNING 186
5.2.3.3 SPINNING DISK CONFOCAL MICROSCOPE 181
5.2.4 CONFOCAL DECONVOLUTION
184
5.3 APPLICATIONS OF CONFOCAL MICROSCOPY 186
5.3.1 NONSCANNING APPLICATIONS 186
5.3.1.1 FLUORESCENCE CORRELATION SPECTROSCOPY 186
5.3.1.2 FLUORESCENCE CROSS-CORRELATION SPECTROSCOPY 190
5.3.1.3 PULSED INTERLEAVED EXCITATION 191
5.3.1.4 BURST ANALYSIS WITH MULTIPARAMETER FLUORESCENCE
DETECTION 193
5.3.2 SCANNING APPLICATIONS BEYOND IMAGING 195
5.3.2.1 NUMBER AND BRIGHTNESS ANALYSIS 195
5.3.2.2 RASTER IMAGE CORRELATION SPECTROSCOPY 198
ACKNOWLEDGMENTS 200
REFERENCES 200
6 TWO-PHOTON EXCITATION MICROSCOPY FOR THREE-DIMENSIONAL
IMAGING OF LIVING INTACT TISSUES 203
DAVID W. PISTON
6.1 INTRODUCTION 203
6.2 WHAT IS TWO-PHOTON EXCITATION? 205
6.2.1 NONLINEAR OPTICS AND 2PM 206
6.2.2 HISTORY AND THEORY OF 2PM 207
6.3 HOW DOES TWO-PHOTON EXCITATION MICROSCOPY WORK IN
PRACTICE? 211
6.3.1 THE ROLE OF LIGHT ABSORPTION IN 2PM 212
6.3.2 THE ROLE OF LIGHT SCATTERING IN 2PM
213
6.4 INSTRUMENTATION 216
6.4.1 LASERS FOR 2PM 216
6.4.2 DETECTION STRATEGIES FOR 2PM 219
6.4.3 THE ADVANTAGES OF 2PM FOR DEEP-TISSUE IMAGING 220
6.5 LIMITATIONS OF TWO-PHOTON EXCITATION MICROSCOPY 222
6.5.1 LIMITS OF SPATIAL RESOLUTION IN 2PM 222
6.5.2 POTENTIAL SAMPLE HEATING BY THE HIGH LASER POWERS IN 2PM 223
6.5.3 DIFFICULTIES IN PREDICTING AND MEASURING TWO-PHOTON EXCITATION
SPECTRA 224
6.5.4 ACCELERATED PHOTOBLEACHING (AND ASSOCIATED PHOTODAMAGE) IN THE
FOCAL PLANE 227
6.5.5 EXPENSIVE LASERS CREATE A PRACTICAL LIMITATION FOR SOME
EXPERIMENTS 228
6.6 WHEN IS 2PM THE BEST OPTION? 229
6.6.1 THICK SPECIMEN INCLUDING
IN VIVO IMAGING 229
6.6.2 IMAGING FLUOROPHORES WITH EXCITATION PEAKS IN THE ULTRAVIOLET (UV)
SPECTRUM 231
6.6.3 LOCALIZED PHOTOCHEMISTRY 231
6.7 APPLICATIONS OF TWO-PHOTON MICROSCOPY 231
6.7.1 IMAGING UV-EXCITED FLUOROPHORES, SUCH AS NADH FOR METABOLIC
ACTIVITY 231
6.7.2 LOCALIZED PHOTOACTIVATION OF CAGED COMPOUNDS 233
6.7.3 IMAGING ELECTRICAL ACTIVITY IN DEEP TISSUE 236
6.7 A LIGHT SHEET MICROSCOPY USING TWO-PHOTON EXCITATION 237
6.7.5 OTHER APPLICATIONS OF 2PM 238
6.8 OTHER NONLINEAR MICROSCOPIES 239
6.9 FUTURE OUTLOOK FOR 2PM 240
6.10 SUMMARY 240
ACKNOWLEDGMENT 241
REFERENCES 241
7 LIGHT SHEET MICROSCOPY 243
GOPI SHAH, MICHAEL WEBER,; AND JAN HUISKEN
7.1 PRINCIPLE OF LIGHT SHEET MICROSCOPY 244
7.2 LIGHT SHEET MICROSCOPY: KEY ADVANTAGES 245
7.3 CONSTRUCTION AND WORKING OF A LIGHT SHEET MICROSCOPE 246
7.4 THEORY OF LIGHT SHEET MICROSCOPY 247
7.5 LIGHT SHEET INTERACTION WITH TISSUE 251
7.6 3D IMAGING 253
171 MULTIVIEW IMAGING 255
7.8 DIFFERENT LENS CONFIGURATIONS 257
7.9 SAMPLE MOUNTING
258
7.10 RECENT ADVANCES IN LIGHT SHEET MICROSCOPY 259
7.11 OUTLOOK
260
7.11.1 BIG DATA 260
7.11.2 SMART MICROSCOPE: IMAGING CONCEPT OF THE FUTURE 261
7.11.3 HIGH-THROUGHPUT IMAGING 261
7.12 SUMMARY 262
REFERENCES 262
8 LOCALIZATION-BASED SUPER-RESOLUTION MICROSCOPY 267
MARKUS SAUER AND MIKE HEILEMANN
8.1 SUPER-RESOLUTION MICROSCOPY: AN INTRODUCTION 267
8.2 THE PRINCIPLE OF SINGLE-MOLECULE LOCALIZATION MICROSCOPY 269
8.3 PHOTOACTIVATABLE AND PHOTOCONVERTIBLE PROBES 272
8.4 INTRINSICALLY PHOTOSWITCHABLE PROBES 272
8.5 PHOTOSWITCHING OF ORGANIC FLUOROPHORES BY CHEMICAL REACTIONS 273
8.6 EXPERIMENTAL SETUP FOR LOCALIZATION MICROSCOPY 273
8.7 OPTICAL RESOLUTION AND IMAGING ARTIFACTS 276
8.8 FLUORESCENCE LABELING FOR SUPER-RESOLUTION MICROSCOPY 278
8.8.1 LABEL SIZE VERSUS STRUCTURAL RESOLUTION 278
8.8.2 LIVE-CELL LABELING 280
8.8.3 CLICK CHEMISTRY 280
8.8.4 THREE-DIMENSIONAL SMLM 281
8.8.5 ASTIGMATIC IMAGING 281
8.8.6 BIPLANE IMAGING 282
8.8.7 DOUBLE HELIX PSF 282
8.8.8 INTERFEROMETRIC IMAGING 282
8.9 MEASURES FOR IMPROVING IMAGING CONTRAST 283
8.10 SMLM SOFTWARE 283
8.11 REFERENCE STRUCTURES FOR SMLM 28S
8.12 QUANTIFICATION OF SMLM DATA 286
8.13 SUMMARY 287
REFERENCES 287
9 SUPER-RESOLUTION MICROSCOPY: INTERFERENCE AND PATTERN
TECHNIQUES 291
UDO BIRK, GERRIT BEST, ROMAN AMBERGER, AND CHRISTOPH CREMER
9.1 INTRODUCTION 291
9.1.1 REVIEW: THE RESOLUTION LIMIT 292
9.2 STRUCTURED ILLUMINATION MICROSCOPY (SIM) 293
9.2.1 IMAGE GENERATION IN STRUCTURED ILLUMINATION MICROSCOPY 295
9.2.2 EXTRACTING THE HIGH-RESOLUTION INFORMATION
298
9.2.3 OPTICAL SECTIONING BY SIM 299
9.2.4 HOW THE ILLUMINATION PATTERN IS GENERATED? 301
9.2.5 MATHEMATICAL DERIVATION OF THE INTERFERENCE PATTERN 302
9.2.6 EXAMPLES FOR SIM SETUPS 304
9.3 SPATIALLY MODULATED ILLUMINATION (SMI) MICROSCOPY 307
9.3.1 OVERVIEW 307
9.3.2 SMI SETUP 309
9.3.3 EXCITATION LIGHT DISTRIBUTION 309
9.3.4 OBJECT SIZE ESTIMATION WITH SMI MICROSCOPY 311
9.4 APPLICATION OF PATTERNED TECHNIQUES 313
9.5 CONCLUSION 317
9.6 SUMMARY 317
ACKNOWLEDGMENTS 317
REFERENCES 318
10 STED MICROSCOPY 321
TRAVIS J. GOULD LENA K. SCHROEDER, PATRINA A. PEL LEFT, AND JOERG
BEWERSDORF
10.1 INTRODUCTION 321
10.2 THE CONCEPTS BEHIND STED MICROSCOPY 322
10.2.1 FUNDAMENTAL CONCEPTS 322
10.2.1.1 SWITCHING BETWEEN OPTICAL STATES 322
10.2.1.2 STIMULATED EMISSION DEPLETION 322
10.2.1.3 STIMULATED EMISSION DEPLETION MICROSCOPY 324
10.2.2 KEY PARAMETERS IN STED MICROSCOPY 326
10.2.2.1 PULSED LASERS AND FLUOROPHORE KINETICS 326
10.2.2.2 WAVELENGTH EFFECTS 328
10.2.2.3 PSF SHAPE AND QUALITY 328
10.3 EXPERIMENTAL SETUP 330
10.3.1 LIGHT SOURCES AND SYNCHRONIZATION 330
10.3.2 SCANNING AND SPEED 331
10.3.3 MULTICOLOR STED IMAGING 332
10.3.4 IMPROVING AXIAL RESOLUTION IN STED MICROSCOPY 333
10.4 APPLICATIONS 334
10.4.1 CHOICE OF FLUOROPHORE 334
10.4.2 LABELING STRATEGIES 335
10.5 SUMMARY 336
REFERENCES 337
11 FLUORESCENCE PHOTOBLEACHING TECHNIQUES 339
REINER PETERS
11.1 INTRODUCTION 339
11.2 BASIC CONCEPTS AND PROCEDURES 340
11.2.1 ONE PRINCIPLE, SEVERAL MODES 340
11.2.2 SETTING UP AN INSTRUMENT 343
11.2.3 APPROACHING COMPLEXITY FROM BOTTOM UP 344
11.3 FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING (FRAP) 345
11.3.1 EVALUATION OF DIFFUSION MEASUREMENTS 345
11.3.2 BINDING 348
11.3.3 MEMBRANE TRANSPORT 349
11.4 CONTINUOUS FLUORESCENCE MICROPHOTOLYSIS (CFM) 352
11.4.1 THEORETICAL BACKGROUND AND DATA EVALUATION 352
11.4.2 COMBINATION OF CFM WITH OTHER TECHNIQUES 355
11.4.3 CFM VARIANTS 355
11.5 CLSM-ASSISTED PHOTOBLEACHING METHODS 356
11.5.1 IMPLEMENTATION 356
11.5.2 NEW OPPORTUNITIES 357
11.5.2.1 MULTIPLE ROPS 357
11.5.2.2 ARBITRARILY SHAPED ROPS 359
11.5.2.3 SPATIALLY RESOLVED BLEACHING AND RECOVERY 359
11.5.2.4 MILLISECOND TIME RESOLUTION 359
11.5.2.5 THREE-DIMENSIONAL PHOTOBLEACHING 359
11.5.3 TWO COMMON ARTIFACTS AND THEIR CORRECTION 360
XII I
CONTENTS
11.6 SUMMARY AND OUTLOOK 360
REFERENCES 361
12 SINGLE-MOLECULE MICROSCOPY IN THE LIFE SCIENCES 365
MARKUS AXMANN, JOSEF MAD I, AND GERHARD J. SCHUETZ
12.1 ENCIRCLING THE PROBLEM 365
12.2 WHAT IS THE UNIQUE INFORMATION? 367
12.2.1 KINETICS CAN BE DIRECTLY RESOLVED 367
12.2.2 FULL PROBABILITY DISTRIBUTIONS CAN BE MEASURED 367
12.2.3 STRUCTURES CAN BE RELATED TO FUNCTIONAL STATES 369
12.2.4 STRUCTURES CAN BE IMAGED AT SUPER-RESOLUTION 370
12.2.5 BIOANALYSIS CAN BE EXTENDED DOWN TO THE SINGLE-MOLECULE LEVEL 372
12.3 BUILDING A SINGLE-MOLECULE MICROSCOPE 372
12.3.1 MICROSCOPES/OBJECTIVES 373
12.3.1.1 DUAL VIEW 373
12.3.1.2 OBJECTIVE 374
12.3.2 LIGHT SOURCE 377
12.3.2.1 UNIFORMITY 377
12.3.2.2 INTENSITY 378
12.3.2.3 ILLUMINATION TIME 380
12.3.2.4 POLARIZATION 380
12.3.2.5 WAVELENGTH 381
12.3.2.6 COLLIMATION 382
12.3.3 DETECTOR 382
12.3.3.1 PIXEL SIZE 382
12.3.3.2 CCD CAMERAS 384
12.3.3.3 ELECTRON-MULTIPLYING CCD CAMERAS 385
12.3.3.4 CMOS DETECTORS 386
12.4 ANALYZING SINGLE-MOLECULE SIGNALS: POSITION, ORIENTATION, COLOR,
AND
BRIGHTNESS 387
12.4.1 LOCALIZING IN TWO DIMENSIONS 388
12.4.2 LOCALIZING ALONG THE OPTICAL AXIS 389
12.4.2.1 ANALYSIS OF THE SHAPE OF THE POINT SPREAD FUNCTION 389
12.4.2.2 INTENSITY PATTERNS ALONG THE OPTICAL AXIS 391
12.4.3 BRIGHTNESS 392
12.4.4 ORIENTATION 392
12.4.4.1 POLARIZATION MICROSCOPY 392
12.4.4.2 DEFOCUSED IMAGING 393
12.4.5 COLOR 393
12.5 LEARNING FROM SINGLE-MOLECULE SIGNALS 394
12.5.1 DETERMINATION OF MOLECULAR ASSOCIATIONS 394
12.5.2 DETERMINATION OF MOLECULAR CONFORMATIONS VIA FRET 395
12.5.3 SINGLE-MOLECULE TRACKING
400
12.5.4 DETECTING TRANSITIONS 401
ACKNOWLEDGMENTS 402
REFERENCES 402
13 FOERSTER RESONANCE ENERGY TRANSFER AND FLUORESCENCE LIFETIME
IMAGING 405
FREDS. WOUTERS
13.1 GENERAL INTRODUCTION 405
13.2 FOERSTER RESONANCE ENERGY TRANSFER 406
13.2.1 PHYSICAL BASIS OF FRET 406
13.2.2 HISTORICAL DEVELOPMENT OF FRET 406
13.2.3 SPECTRAL AND DISTANCE DEPENDENCE OF FRET 416
13.2.4 FRET IS OF LIMITED USE AS A MOLECULAR RULER 420
13.2.5 SPECIAL FRET CONDITIONS 422
13.2.5.1 DIFFUSION-ENHANCED FRET 422
13.2.5.2 MULTIPLE ACCEPTORS 422
13.2.5.3 FRET IN A PLANE 423
13.3 MEASURING FRET 426
13.3.1 SPECTRAL CHANGES 426
13.3.1.1 FRET FROM DONOR QUENCHING 426
13.3.1.2 FRET-INDUCED ACCEPTOR EMISSION 427
13.3.1.3 CONTRAST IN INTENSITY-BASED FRET MEASUREMENTS 430
13.3.1.4 FULL QUANTITATION OF INTENSITY-BASED FRET MEASUREMENTS 431
13.3.1.5 OCCUPANCY ERRORS IN FRET 432
13.3.2 DECAY KINETICS 432
13.3.2.1 PHOTOBLEACHING RATE 432
13.3.2.2 FLUORESCENCE LIFETIME CHANGES 435
13.4 FLIM 439
13.4.1 FREQUENCY-DOMAIN FLIM 441
13.4.1.1 OPERATION PRINCIPLE AND TECHNICAL ASPECTS 441
13.4.2 TIME-DOMAIN FLIM 442
13.4.2.1 TIME-CORRELATED SINGLE-PHOTON COUNTING 442
13.4.2.2 TIME GATING 443
13.5 ANALYSIS AND PITFALLS 444
13.5.1 AVERAGE LIFETIME, MULTIPLE LIFETIME FITTING 444
13.5.2 FROM FRET/LIFETIME TO SPECIES 444
13.6 SUMMARY 448
REFERENCES 448
A APPENDIX A: WHAT EXACTLY IS A DIGITAL IMAGE? 453
ULRICH KUBITSCHECK
A.L INTRODUCTION 453
A.2 DIGITAL IMAGES AS MATRICES 453
A.2.1 GRAY VALUES AS A FUNCTION OF SPACE AND TIME 453
A.2.1.1 PARALLEL DATA ACQUISITION 454
A.2.1.2 SEQUENTIAL DATA ACQUISITION 455
A.2.2 IMAGE SIZE, BIT DEPTH, AND STORAGE REQUIREMENTS 456
A.3 LOOK-UP TABLE 457
A.4 INTENSITY HISTOGRAMS 457
A.5 IMAGE PROCESSING 458
A.5.1 OPERATIONS ON SINGLE PIXELS 458
A.5.2
OPERATIONS ON PIXEL GROUPS 459
A.5.3 LOW-PASS FILTERS 460
A.6
PITFALLS 460
A.7 SUMMARY 461
REFERENCES 461
B
APPENDIX B: PRACTICAL GUIDE TO OPTICAL ALIGNMENT
463
RAINER HEINTZMANN
B.L HOW TO OBTAIN A WIDENED PARALLEL LASER BEAM?
463
B.2 MIRROR ALIGNMENT 465
B.3 LENS ALIGNMENT 466
B.4 AUTOCOLLIMATION TELESCOPE 466
B.5 ALIGNING A SINGLE LENS USING A LASER BEAM
466
B.6 HOW TO FIND THE FOCAL PLANE OF A LENS? 469
B.7 HOW TO FOCUS TO THE BACK FOCAL PLANE OF AN OBJECTIVE LENS? 470
INDEX 473
|
| any_adam_object | 1 |
| author2 | Kubitscheck, Ulrich |
| author2_role | edt |
| author2_variant | u k uk |
| author_GND | (DE-588)1131264150 |
| author_facet | Kubitscheck, Ulrich |
| building | Verbundindex |
| bvnumber | BV043811130 |
| classification_rvk | UH 5870 UH 6700 WC 2900 WC 2905 |
| classification_tum | CHE 243f CHE 264f |
| ctrlnum | (OCoLC)989949893 (DE-599)BVBBV043811130 |
| dewey-full | 502.82 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 502 - Miscellany |
| dewey-raw | 502.82 |
| dewey-search | 502.82 |
| dewey-sort | 3502.82 |
| dewey-tens | 500 - Natural sciences and mathematics |
| discipline | Allgemeine Naturwissenschaft Physik Biologie Chemie |
| edition | Second edition |
| format | Book |
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| id | DE-604.BV043811130 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T07:35:42Z |
| institution | BVB |
| isbn | 9783527338375 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-029222383 |
| oclc_num | 989949893 |
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| owner_facet | DE-29T DE-11 DE-355 DE-BY-UBR DE-20 DE-384 DE-523 |
| physical | xxii, 482 Seiten Illustrationen, Diagramme (farbig) |
| publishDate | 2017 |
| publishDateSearch | 2017 |
| publishDateSort | 2017 |
| publisher | Wiley-VCH |
| record_format | marc |
| spelling | Fluorescence microscopy from principles to biological applications edited by Ulrich Kubitscheck Second edition Weinheim Wiley-VCH [2017] © 2017 xxii, 482 Seiten Illustrationen, Diagramme (farbig) txt rdacontent n rdamedia nc rdacarrier Fluoreszenzmikroskopie (DE-588)4290958-2 gnd rswk-swf 1\p (DE-588)1071861417 Konferenzschrift gnd-content Fluoreszenzmikroskopie (DE-588)4290958-2 s DE-604 Kubitscheck, Ulrich (DE-588)1131264150 edt Erscheint auch als Online-Ausgabe, ePDF 978-3-527-68772-5 Erscheint auch als Online-Ausgabe, ePub 978-3-527-68774-9 Erscheint auch als Online-Ausgabe, Mobi 978-3-527-68775-6 Erscheint auch als Online-Ausgabe, oBook 978-3-527-68773-2 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029222383&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Fluorescence microscopy from principles to biological applications Fluoreszenzmikroskopie (DE-588)4290958-2 gnd |
| subject_GND | (DE-588)4290958-2 (DE-588)1071861417 |
| title | Fluorescence microscopy from principles to biological applications |
| title_auth | Fluorescence microscopy from principles to biological applications |
| title_exact_search | Fluorescence microscopy from principles to biological applications |
| title_full | Fluorescence microscopy from principles to biological applications edited by Ulrich Kubitscheck |
| title_fullStr | Fluorescence microscopy from principles to biological applications edited by Ulrich Kubitscheck |
| title_full_unstemmed | Fluorescence microscopy from principles to biological applications edited by Ulrich Kubitscheck |
| title_short | Fluorescence microscopy |
| title_sort | fluorescence microscopy from principles to biological applications |
| title_sub | from principles to biological applications |
| topic | Fluoreszenzmikroskopie (DE-588)4290958-2 gnd |
| topic_facet | Fluoreszenzmikroskopie Konferenzschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029222383&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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