Fluorescence microscopy: from principles to biological applications
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Weinheim
Wiley-Blackwell
2013
|
| Subjects: | |
| Online Access: | Inhaltstext Inhaltsverzeichnis |
| Physical Description: | XX, 410 S. Ill., graph. Darst. |
| ISBN: | 3527329226 9783527671601 9783527671618 9783527671625 9783527671595 9783527329229 |
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IMAGE 1
CONTENTS
PREFACE XIII LIST O F CONTRIBUTORS XVII
1 INTRODUCTION TO OPTICS AND PHOTOPHYSICS 1
RAINER HEINTZMANN
1.1 INTERFERENCE: LIGHT AS A WAVE 2
1.2 TWO EFFECTS O F INTERFERENCE: DIFFRACTION AND REFRACTION 7 1.3
OPTICAL ELEMENTS 14
1.3.1 LENSES 14
1.3.2 METALLIC MIRROR 17
1.3.3 DIELECTRIC MIRROR 18
1.3.4 PINHOLES 18
1.3.5 FILTERS 19
1.3.6 CHROMATIC REFLECTORS 20
1.4 THE FAR-FIELD, NEAR-FIELD, AND EVANESCENT WAVES 20 1.5 OPTICAL
ABERRATIONS 23
1.6 PHYSICAL BACKGROUND O F FLUORESCENCE 24 1.7 PHOTONS, POISSON
STATISTICS, AND ANTIBUNCHING 30 REFERENCES 31
2 PRINCIPLES O F LIGHT MICROSCOPY 33
ULRICH KUBITSCHECK
2.1 INTRODUCTION 33
2.2 CONSTRUCTION O F LIGHT MICROSCOPES 33
2.2.1 COMPONENTS OF LIGHT MICROSCOPES 33 2.2.2 IMAGING PATH 34
2.2.3 MAGNIFICATION 36
2.2.4 ANGULAR AND NUMERICAL APERTURE 38 2.2.5 FIELD O F VIEW 38
2.2.6 ILLUMINATION BEAM PATH 39
2.3 WAVE OPTICS AND RESOLUTION 42
2.3.1 WAVE OPTICAL DESCRIPTION O F THE IMAGING PROCESS 43
HTTP://D-NB.INFO/1027115853
IMAGE 2
V I I CONTENTS
2.3.2 THE AIRY FUNCTION 47
2.3.3 POINT SPREAD FUNCTION AND OPTICAL TRANSFER FUNCTION 50 2.3.4
LATERAL AND AXIAL RESOLUTION 52 2.3.5 MAGNIFICATION AND RESOLUTION 59
2.3.6 DEPTH O F FIELD AND DEPTH OF FOCUS 60
2.3.7 OVER- AND UNDER SAMPLING 61
2.4 APERTURES, PUPILS, AND TELECENTRICITY 61 2.5 MICROSCOPE OBJECTIVES
64
2.5.1 OBJECTIVE LENS DESIGN 64
2.5.2 LIGHT COLLECTION EFFICIENCY AND IMAGE BRIGHTNESS 68 2.5.3
OBJECTIVE LENS CLASSES 73
2.5.4 IMMERSION MEDIA 73
2.5.5 SPECIAL APPLICATIONS 77
2.6 CONTRAST 78
2.6.1 DARK FIELD 80
2.6.2 PHASE CONTRAST 81
2.6.3 INTERFERENCE CONTRAST 86
2.6.4 ADVANCED TOPIC: DIFFERENTIAL INTERFERENCE CONTRAST 89 2.7 SUMMARY
94
ACKNOWLEDGMENTS 94 REFERENCES 95
3 FLUORESCENCE MICROSCOPY 97
JUREK W. DOBRUCKI
3.1 FEATURES O F FLUORESCENCE MICROSCOPY 98 3.1.1 * IMAGE CONTRAST 98
3.1.2 SPECIFICITY OF FLUORESCENCE LABELING 101 3.1.3 SENSITIVITY OF
DETECTION 102 3.2 A FLUORESCENCE MICROSCOPE 103
3.2.1 PRINCIPLE OF OPERATION 103
3.2.2 SOURCES O F EXCITING LIGHT 107 3.2.3 OPTICAL FILTERS IN A
FLUORESCENCE MICROSCOPE 110 3.2.4 ELECTRONIC FILTERS 111
3.2.5 PHOTODETECTORS FOR FLUORESCENCE MICROSCOPY 112 3.2.6
CCD-CHARGE-COUPLED DEVICE 113 3.2.7 INTENSIFIED CCD (ICCD) 116 3.2.8
ELECTRON-MULTIPLYING CHARGE-COUPLED DEVICE (EMCCD) 117
3.2.9 CMOS 119
3.2.10 SCIENTIFIC CMOS (SCMOS) 120 3.2.11 FEATURES O F CCD AND CMOS
CAMERAS 121 3.2.12 CHOOSING A DIGITAL CAMERA FOR FLUORESCENCE MICROSCOPY
121 3.2.13 PHOTOMULTIPLIER TUBE (PMT) 121 3.2.14 AVALANCHE PHOTODIODE
(APD) 122
IMAGE 3
CONTENTS I V I I
3.3 TYPES O F NOISE IN A DIGITAL MICROSCOPY IMAGE 123
3.4 QUANTITATIVE FLUORESCENCE MICROSCOPY 127 3.4.1 MEASUREMENTS O F
FLUORESCENCE INTENSITY AND CONCENTRATION O F THE LABELED TARGET 127
3.4.2 RATIOMETRIC MEASUREMENTS (CA ++ , PH) 130 3.4.3 MEASUREMENTS OF
DIMENSIONS IN 3D FLUORESCENCE
MICROSCOPY 131
3.4.4 MEASUREMENTS O F EXCITING LIGHT INTENSITY 132 3.4.5 TECHNICAL TIPS
FOR QUANTITATIVE FLUORESCENCE MICROSCOPY 132 3.5 LIMITATIONS O F
FLUORESCENCE MICROSCOPY 133 3.5.1 PHOTOBLEACHING 133
3.5.2 REVERSIBLE PHOTOBLEACHING UNDER OXIDIZING OR REDUCING CONDITIONS
134 3.5.3 PHOTOTOXICITY 135
3.5.4 OPTICAL RESOLUTION 135
3.5.5 MISREPRESENTATION O F SMALL OBJECTS 3.6 CURRENT AVENUES OF
DEVELOPMENT REFERENCES 139 FURTHER READING 141
RECOMMENDED INTERNET RESOURCES FLUORESCENT SPECTRA DATABASE 142
4 FLUORESCENCE LABELING 143
GERD ULRICH NIENHAUS AND KARIN NIENHAUS 4.1 INTRODUCTION 143
4.2 PRINCIPLES O F FLUORESCENCE 143
4.3 KEY PROPERTIES O F FLUORESCENT LABELS 144 4.4 SYNTHETIC FLUOROPHORES
149
4.4.1 ORGANIC DYES 149
4.4.2 FLUORESCENT NANOPARTICLES 150 4.4.3 CONJUGATION STRATEGIES FOR
SYNTHETIC FLUOROPHORES 152 4.4.4 NONNATURAL AMINO ACIDS 155
4.4.5 BRINGING THE FLUOROPHORE TO ITS TARGET 156 4.5 GENETICALLY ENCODED
LABELS 158
4.5.1 PHYCOBILIPROTEINS 158
4.5.2 GFP-LIKE PROTEINS 159
4.6 LABEL SELECTION FOR PARTICULAR APPLICATIONS 163 4.6.1 FRET TO
MONITOR INTRAMOLECULAR CONFORMATIONAL DYNAMICS 163 4.6.2 PROTEIN
EXPRESSION IN CELLS 167
4.6.3 FLUOROPHORES AS SENSORS INSIDE THE CELL 167 4.6.4 LIVE-CELL
DYNAMICS 168
4.7 CONCLUSIONS 168
REFERENCES 169
137 139
142
IMAGE 4
V I I I I CONTENTS
5 CONFOCAL MICROSCOPY 175
NIKOLAUS NAREDI-RAINER, JENS PRESCHER, ACHIM HARTSCHUH, AND DON C. LAMB
5.1 INTRODUCTION 175
5.1.1 EVOLUTION AND LIMITS O F CONVENTIONAL WIDEFIELD MICROSCOPY 175
5.1.2 HISTORY AND DEVELOPMENT OF CONFOCAL MICROSCOPY 177 5.2 THE THEORY
O F CONFOCAL MICROSCOPY 180
5.2.1 THE PRINCIPLE O F CONFOCAL MICROSCOPY 180 5.2.2 RADIAL AND AXIAL
RESOLUTION AND THE IMPACT O F THE PINHOLE SIZE 182 5.2.3 SCANNING
CONFOCAL IMAGING 189
5.2.4 CONFOCAL DECONVOLUTION 194 5.3 APPLICATIONS OF CONFOCAL MICROSCOPY
196 5.3.1 NONSCANNING APPLICATIONS 196 5.3.2 ADVANCED CORRELATION
TECHNIQUES 200
5.3.3 SCANNING APPLICATIONS BEYOND IMAGING 205 ACKNOWLEDGMENTS 210
REFERENCES 210
6 FLUORESCENCE PHOTOBLEACHING AND PHOTOACTIVATION TECHNIQUES 215 REINER
PETERS 6.1 INTRODUCTION 215
6.2 BASIC CONCEPTS AND PROCEDURES 236
6.2.1 PUTTING PHOTOBLEACHING TO WORK 216 6.2.2 SETTING UP AN INSTRUMENT
219
6.2.3 APPROACHING COMPLEXITY FROM BOTTOM UP 220 6.3 FLUORESCENCE
RECOVERY AFTER PHOTOBLEACHING (FRAP) 221 6.3.1 EVALUATION O F DIFFUSION
MEASUREMENTS 222 6.3.2 BINDING 225
6.3.3 MEMBRANE TRANSPORT 226
6.4 CONTINUOUS FLUORESCENCE MICROPHOTOLYSIS (CFM) 228 6.4.1 THEORETICAL
BACKGROUND AND DATA EVALUATION 229 6.4.2 COMBINATION O F CFM WITH OTHER
TECHNIQUES 231 6.4.3 CFM VARIANTS 232
6.5 CONFOCAL PHOTOBLEACHING 233
6.5.1 USE O F LASER SCANNING MICROSCOPES (LSMS) IN PHOTOBLEACHING
EXPERIMENTS 233 6.5.2 NEW POSSIBILITIES PROVIDED BY CONFOCAL
PHOTOBLEACHING 234 6.5.3 ARTIFACTS AND REMEDIES 237
6.6 FLUORESCENCE PHOTOACTIVATION AND DISSIPATION 238 6.6.1 BASIC ASPECTS
238
6.6.2 THEORY AND INSTRUMENTATION 239 6.6.3 REVERSIBLE FLUX MEASUREMENTS
239
IMAGE 5
CONTENTS IIX
6.7 SUMMARY AND OUTLOOK 241
REFERENCES 241
7 FDRSTER RESONANCE ENERGY TRANSFER AND FLUORESCENCE LIFETIME IMAGING
245 FRED S. WOUTERS
7.1 GENERAL INTRODUCTION 245
7.2 FRET 246
7.2.1 HISTORICAL DEVELOPMENT O F FRET 246 7.2.2 REQUIREMENTS 254
7.2.3 FRET AS A MOLECULAR RULER 258
7.2.4 SPECIAL FRET CONDITIONS 262 7.3 MEASURING FRET 265
7.3.1 SPECTRAL CHANGES 266
7.3.2 DECAY KINETICS 272
7.4 FLIM 280
7.4.1 FREQUENCY-DOMAIN FLIM 282 7.4.2 TIME-DOMAIN FLIM 283
7.5 ANALYSIS AND PITFALLS 285
7.5.1 AVERAGE LIFETIME, MULTIPLE LIFETIME FITTING 285 7.5.2 FROM
FRET/LIFETIME TO SPECIES 286 SUMMARY 287 REFERENCES 288
8 SINGLE-MOLECULE MICROSCOPY IN THE LIFE SCIENCES 293 MARKUS AXMANN,
JOSEF MADL, AND GERHARD J. SCHIITZ 8.1 ENCIRCLING THE PROBLEM 293
8.2 WHAT IS THE UNIQUE INFORMATION? 295
8.2.1 KINETICS CAN BE DIRECTLY RESOLVED 295 8.2.2 FULL PROBABILITY
DISTRIBUTIONS CAN BE MEASURED 296 8.2.3 STRUCTURES CAN BE RELATED TO
FUNCTIONAL STATES 297 8.2.4 STRUCTURES CAN BE IMAGED AT SUPERRESOLUTION
298 8.2.5 BIOANALYSIS CAN BE EXTENDED DOWN
TO THE SINGLE-MOLECULE LEVEL 300 8.3 BUILDING A SINGLE-MOLECULE
MICROSCOPE 301 8.3.1 MICROSCOPES/OBJECTIVES 301 8.3.2 LIGHT SOURCE 304
8.3.3 DETECTOR 310
8.4 ANALYZING SINGLE-MOLECULE SIGNALS: POSITION, ORIENTATION, COLOR, AND
BRIGHTNESS 316 8.4.1 LOCALIZING IN TWO DIMENSIONS 316 8.4.2 LOCALIZING
ALONG THE OPTICAL AXIS 318
8.4.3 BRIGHTNESS 320
8.4.4 ORIENTATION 321
8.4.5 COLOR 322
IMAGE 6
X CONTENTS
8.5 LEARNING FROM SINGLE-MOLECULE SIGNALS 323
8.5.1 DETERMINATION OF MOLECULAR ASSOCIATIONS 323 8.5.2 DETERMINATION OF
MOLECULAR CONFORMATIONS VIA FRET 325 8.5.3 SUPERRESOLUTION
SINGLE-MOLECULE MICROSCOPY 329 8.5.4 SINGLE-MOLECULE TRACKING 332 8.5.5
DETECTING TRANSITIONS 332
ACKNOWLEDGMENTS 334 REFERENCES 334
9 SUPER-RESOLUTION MICROSCOPY: INTERFERENCE AND PATTERN TECHNIQUES 345
GERRIT BEST, ROMAN AMBERGER, AND CHRISTOPH CREMER 9.1 INTRODUCTION 345
9.1.1 REVIEW: THE RESOLUTION LIMIT 346 9.2 STRUCTURED ILLUMINATION
MICROSCOPY (SIM) 347 9.2.1 IMAGE GENERATION IN STRUCTURED ILLUMINATION
MICROSCOPY 349 9.2.2 EXTRACTING THE HIGH-RESOLUTION INFORMATION 352
9.2.3 OPTICAL SECTIONING BY SIM 353 9.2.4 HOW THE ILLUMINATION PATTERN
IS GENERATED 355 9.2.5 MATHEMATICAL DERIVATION OF THE INTERFERENCE
PATTERN 355 9.2.6 EXAMPLES FOR SIM SETUPS 358
9.3 SPATIALLY MODULATED ILLUMINATION (SMI) MICROSCOPY 362 9.3.1 OVERVIEW
362
9.3.2 SMI SETUP 363
9.3.3 THE OPTICAL PATH 364
9.3.4 SIZE ESTIMATION WITH SMI MICROSCOPY 366 9.4 APPLICATION OF
PATTERNED TECHNIQUES 368 9.5 CONCLUSION 372
9.6 SUMMARY 372
ACKNOWLEDGMENTS 373 REFERENCES 373
10 STED MICROSCOPY 375
TRAVIS J. GOULD, PATRINA A. PELLETT, AND JOERG BEWERSDORF 10.1
INTRODUCTION 375
10.2 THE CONCEPTS BEHIND STED MICROSCOPY 376 10.2.1 FUNDAMENTAL CONCEPTS
376 10.2.2 KEY PARAMETERS IN STED MICROSCOPY 380 10.3 EXPERIMENTAL SETUP
384
10.3.1 LIGHT SOURCES AND SYNCHRONIZATION 384 10.3.2 SCANNING AND SPEED
385
10.3.3 MULTICOLOR STED IMAGING 386 10.3.4 IMPROVING AXIAL RESOLUTION IN
STED MICROSCOPY 388 10.4 APPLICATIONS 388
10.4.1 CHOICE OF FLUOROPHORE 388
IMAGE 7
CONTENTS X I
10.4.2 LABELING STRATEGIES 389
SUMMARY 390 REFERENCES 391
A APPENDIX: PRACTICAL GUIDE TO OPTICAL ALIGNMENT 393 RAINER HEINTZMANN
A.L HOW TO OBTAIN A WIDENED PARALLEL LASER BEAM 393 A.2 MIRROR ALIGNMENT
395
A.3 LENS ALIGNMENT 396
A.4 AUTOCOLLIMATION TELESCOPE 396 A.5 ALIGNING A SINGLE LENS USING A
LASER BEAM 397 A.6 HOW TO FIND THE FOCAL PLANE O F A LENS 399
A.7 HOW TO FOCUS TO THE BACK FOCAL PLANE O F AN OBJECTIVE LENS 400
INDEX 403 |
| any_adam_object | 1 |
| author_GND | (DE-588)1131264150 |
| building | Verbundindex |
| bvnumber | BV040774845 |
| classification_rvk | UH 5870 UH 6700 WC 2900 WC 2905 |
| classification_tum | CHE 264f CHE 243f |
| ctrlnum | (OCoLC)843434348 (DE-599)DNB1027115853 |
| 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 |
| format | Book |
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| genre | 1\p (DE-588)1071861417 Konferenzschrift gnd-content |
| genre_facet | Konferenzschrift |
| id | DE-604.BV040774845 |
| illustrated | Illustrated |
| indexdate | 2024-08-21T00:34:10Z |
| institution | BVB |
| isbn | 3527329226 9783527671601 9783527671618 9783527671625 9783527671595 9783527329229 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025753184 |
| oclc_num | 843434348 |
| open_access_boolean | |
| owner | DE-11 DE-91G DE-BY-TUM DE-20 DE-703 DE-19 DE-BY-UBM DE-83 DE-91 DE-BY-TUM DE-355 DE-BY-UBR DE-29T |
| owner_facet | DE-11 DE-91G DE-BY-TUM DE-20 DE-703 DE-19 DE-BY-UBM DE-83 DE-91 DE-BY-TUM DE-355 DE-BY-UBR DE-29T |
| physical | XX, 410 S. Ill., graph. Darst. |
| publishDate | 2013 |
| publishDateSearch | 2013 |
| publishDateSort | 2013 |
| publisher | Wiley-Blackwell |
| record_format | marc |
| spelling | Fluorescence microscopy from principles to biological applications ed. by Ulrich Kubitscheck Weinheim Wiley-Blackwell 2013 XX, 410 S. Ill., graph. Darst. 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 Sonstige (DE-588)1131264150 oth Erscheint auch als Online-Ausgabe Fluorescence microscopy (DE-604)BV042341295 text/html http://deposit.dnb.de/cgi-bin/dokserv?id=4166015&prov=M&dok_var=1&dok_ext=htm Inhaltstext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025753184&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 ed. by Ulrich Kubitscheck |
| title_fullStr | Fluorescence microscopy from principles to biological applications ed. by Ulrich Kubitscheck |
| title_full_unstemmed | Fluorescence microscopy from principles to biological applications ed. 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://deposit.dnb.de/cgi-bin/dokserv?id=4166015&prov=M&dok_var=1&dok_ext=htm http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025753184&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT kubitscheckulrich fluorescencemicroscopyfromprinciplestobiologicalapplications |