Application of nanoparticles in correlative super-resolution light and electron microscopy:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2014
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 131 Bl. Ill., graph. Darst. 30 cm |
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| adam_text | APPLICATION OF NANOPARTICLES IN CORRELATIVE SUPER-RESOLUTION
LIGHT AND ELECTRON MICROSCOPY
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DISSERTATION
ZUR ERLANGUNG DER WURDE DES DOKTORS DER NATURWISSENSCHAFTEN
AN DER FAKULTAT FUR MATHEMATIK, INFORMATIK UND NATURWISSENSCHAFTEN
FACHBEREICH CHEMIE
DER UNIVERSITAT HAMBURG
VORGELEGT VON
DENNIS EGGERT
HAMBURG, 2014
HTTP://D-NB.INFO/1054708045
TABLE OF CONTENTS
TABLE OF CONTENTS
1. INTRODUCTION 1
1.1. FLUORESCENCE LIGHT MICROSCOPY AND ITS LIMITS 1
1.2. RESOLUTION IN LIGHT MICROSCOPY 1
1.3. SUPER-RESOLUTION MICROSCOPY 3
1.3.1. STOCHASTIC OPTICAL RECONSTRUCTION MICROSCOPY 4
1.3.1.1. THE PRINCIPLE OF STORM 4
1.3.1.2. DYES
FOR STORM 7
1.3.1.3. MULTICOLOR STORM 8
1.3.1.4. DIRECT STOCHASTIC OPTICAL RECONSTRUCTION MICROSCOPY (DSTORM) 10
1.3.2. IMPROVEMENTS IN LOCALIZATION MICROSCOPY 10
1.3.3. APPLICATION OF SUPER-RESOLUTION MICROSCOPY IN CELL BIOLOGY 11
1.4. CORRELATIVE MICROSCOPY 11
1.5. NANOPARTICLES IN BIOIMAGING 13
2. AIM
OF THE THESIS 15
3. MATERIALS AND METHODS 17
3.1. CULTURE OF HUMAN CELLS 17
3.2. LONG
TERM STORAGE OF CELLS 17
3.3. CULTURE OF HUMAN CYTOMEGALOVIRUSES (HCMV) 17
3.4. LABELING OF ANTIBODIES FOR STORM 18
3.5. INDIRECT IMMUNOCYTOCHEMICAL STAINING 19
3.6. CYTOSKELETON EXTRACTION 22
3.7. PREPARATION OF TOKUYASU CROYSECTIONS 23
3.7.1. COATING OF COVERSLIPS WITH ALCIAN BLUE 23
3.7.2. IMMUNOIABELING OF TOKUYASU CRYOSECTIONS 24
3.8.
PREPARATION OF IMMUNOGOLD-REPLICAS 25
3.9. SUPER-RESOLUTION MICROSCOPY 25
3.9.1. BUFFER COMPOSITIONS FOR SUPER-RESOLUTION MICROSCOPY 25
3.9.2. MICROSCOPE SETUP 26
3.9.3. IMAGE ACQUISITION 27
3.9.3.1. STORM 27
3.9.3.2. DSTORM,
PALM AND LOCALIZATION MICROSCOPY 28
3.9.4. IMAGE COMPUTATION 29
3.9.4.1. STORM 29
TABLE OF CONTENTS
3.9.4.2. DSTORM,
PALM AND LOCALIZATION MICROSCOPY 30
3.9.4.3. BAYESIAN LOCALIZATION MICROSCOPY 31
3.9.5. DRIFT CORRECTION 31
3.10. PHASE CONTRAST MICROSCOPY 31
3.10.1. SHADING
CORRECTION 32
3.11. DARKFIELD MICROSCOPY 32
3.12. CONFOCAL LASER SCANNING
MICROSCOPY (CLSM) 32
3.13. HIGH RESOLUTION DIC IMAGING 33
3.14. 3D VISUALIZATION OF MICROSCOPIC IMAGES 33
3.15. TRANSMISSION ELECTRON MICROSCOPY 33
3.16. IMAGE ANALYSIS AND
STATISTICAL ANALYSIS 33
3.17. SPECTROSCOPIC ANALYSIS OF NANOPARTICLES 34
3.18. LABELING OF INFLUENZA A VIRUS PARTICLES WITH AMINE REACTIVE DYES
34
3.18.1. FLUORESCENCE MICROSCOPY 35
3.18.2. IMAGE PROCESSING 35
3.19. CORRELATION OF LIGHT AND ELECTRON MICROSCOPIC IMAGES 35
4. RESULTS 36
4.1. DIFFERENT APPROACHES OF SUPER-RESOLUTION MICROSCOPY ON A COMMERCIAL
STORM
MICROSCOPE 36
4.1.1. EVALUATION OF OPTIMAL CONDITIONS FOR SINGLE COLOR, MULTICOLOR AND
3D
STORM 37
4.1.2. ESTABLISHMENT OF OPTIMAL CONDITIONS FOR DSTORM ON THE NIKON
N-STORM
MICROSCOPE 40
4.1.3. COMBINED DSTORM/PALM IMAGING 48
4.1.4. APPLICATION OF LOCALIZATION MICROSCOPY IN PLANT ORGANS 49
4.2. HIGH RESOLUTION TRANSMITTED LIGHT MICROSCOPY 53
4.2.1. VISUALIZATION OF CELL PERMEABILIZATION ARTIFACTS BY HIGH
RESOLUTION
TRANSMITTED LIGHT MICROSCOPY 57
4.3. (D)STORM USING
GLASS COVERSLIPS 59
4.4. CORRELATION OF SUPER-RESOLUTION FLUORESCENCE WITH HIGH RESOLUTION
TRANSMITTED LIGHT
MICROSCOPY 62
4.5. APPLICATION OF COMPOSITE NANOPARTICLES IN MICROSCOPY 65
4.5.1. COMPOSITE NANOPARTICLES AS UNIVERSAL FIDUCIAL MARKERS 65
4.5.2. CORRELATION OF (D)STORM WITH HIGH RESOLUTION PHASE CONTRAST
MICROSCOPY
USING
COMPOSITE NANOPARTICLES AS FIDUCIAL MARKERS 66
TABLE OF CONTENTS
4.5.3. DRIFT CORRECTION USING NANOPARTICLE-BASED FIDUCIAL MARKERS 68
4.5.4. COMPOSITE NANOPARTICLES AS A SOLID STATE REFERENCE OBJECT FOR
(D)STORM 74
4.6. NANOPARTICLES AS DYES FOR
SINGLE MOLECULE LOCALIZATION MICROSCOPY 77
4.6.1. ANALYSIS OF THE BLINKING OF QUANTUM DOTS 77
4.6.2. QUANTUM DOTS AS DYES FOR (D)STORM 81
4.6.3. QUANTUM DOTS AS DYES FOR BAYESIAN LOCALIZATION MICROSCOPY 83
4.7. CORRELATIVE LIGHT AND ELECTRON MICROSCOPY (CLEM) 86
4.7.1. CORRELATIVE FLUORESCENCE LIGHT AND TRANSMISSION ELECTRON
MICROSCOPY OF
FIDUCIAL-LIKE OBJECTS 86
4.7.2.
CORRELATION OF SUPER-RESOLUTION FLUORESCENCE LIGHT AND TRANSMISSION
ELECTRON
MICROSCOPY USING NANOPARTICLES 88
5. DISCUSSION 92
5.1. COMPARISON OF DIFFERENT DYES FOR STORM,
DSTORM AND LOCALIZATION
MICROSCOPY 93
5.2. APPLICATION OF COMPOSITE NANOPARTICLES IN MICROSCOPY 97
5.3. CORRELATION OF (D)STORM AND
HIGH RESOLUTION TRANSMITTED LIGHT MICROSCOPY 99
5.4. CORRELATION OF (D)STORM AND
TRANSMISSION ELECTRON MICROSCOPY 100
6. ZUSAMMENFASSUNG 102
7. SUMMARY 104
8. LITERATURE 106
9. APPENDIX 113
9.1. ABBREVIATIONS 113
9.2. LIST OF FIGURES 116
9.3.
LIST OF TABLES 118
9.4. LIST OF HAZARDOUS CHEMICALS 119
9.4.1. GHS
HAZARD PICTOGRAMS 122
9.4.2. GHS HAZARD
STATEMENTS 122
9.4.3. GHS
PRECAUTIONARY STATEMENTS 124
9.5. PRIZES,
PUBLICATIONS AND PRESENTATIONS IN CONTEXT OF THE DOCTORAL THESIS 127
9.6. CURRICULUM VITAE 129
9.7. ACKNOWLEDGEMENTS 130
9.8. EIDESSTATTLICHE VERSICHERUNG 131
|
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| author_facet | Eggert, Dennis |
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| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 570 - Biology |
| dewey-raw | 570.282 |
| dewey-search | 570.282 |
| dewey-sort | 3570.282 |
| dewey-tens | 570 - Biology |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| spellingShingle | Eggert, Dennis Application of nanoparticles in correlative super-resolution light and electron microscopy Hochauflösendes Verfahren (DE-588)4287503-1 gnd Elektronenmikroskopie (DE-588)4014327-2 gnd Nanopartikel (DE-588)4333369-2 gnd Fluoreszenzmikroskopie (DE-588)4290958-2 gnd |
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| title | Application of nanoparticles in correlative super-resolution light and electron microscopy |
| title_auth | Application of nanoparticles in correlative super-resolution light and electron microscopy |
| title_exact_search | Application of nanoparticles in correlative super-resolution light and electron microscopy |
| title_full | Application of nanoparticles in correlative super-resolution light and electron microscopy vorgelegt von Dennis Eggert |
| title_fullStr | Application of nanoparticles in correlative super-resolution light and electron microscopy vorgelegt von Dennis Eggert |
| title_full_unstemmed | Application of nanoparticles in correlative super-resolution light and electron microscopy vorgelegt von Dennis Eggert |
| title_short | Application of nanoparticles in correlative super-resolution light and electron microscopy |
| title_sort | application of nanoparticles in correlative super resolution light and electron microscopy |
| topic | Hochauflösendes Verfahren (DE-588)4287503-1 gnd Elektronenmikroskopie (DE-588)4014327-2 gnd Nanopartikel (DE-588)4333369-2 gnd Fluoreszenzmikroskopie (DE-588)4290958-2 gnd |
| topic_facet | Hochauflösendes Verfahren Elektronenmikroskopie Nanopartikel Fluoreszenzmikroskopie Hochschulschrift |
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