Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Online Access: | Volltext Inhaltsverzeichnis |
| Item Description: | Zsfassung in dt. und engl. Sprache |
| Physical Description: | 86 S. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells |c von Meike Heidbreder |
| 246 | 1 | 3 | |a Assoziierung und Aktivierung des TNF-Rezeptor I untersucht mit Einzelmolekül-Tracking und hochauflösender Mikroskopie in lebenden Zellen |
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Record in the Search Index
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| adam_text | IMAGE 1
T A B L E O F C O N T E N T S
SUMMARY 1
ZUSAMMENFASSUNG 3
1. INTRODUCTION 1
1.1. TNF ALPHA 2
1.2. THE TNF SUPERFAMILY - STRUCTURE 3
1.2.1. SIGNALLING VIA TNF-R1 / TNF-R2 4
1.2.2. THE ROLE O F LIPID RAFTS 6
1.2.3. TNF-R PRE-ASSEMBLY 8
1.3. AIMS O F THIS WORK 9
2. THEORETICAL BACKGROUND 10
2.1. FLUORESCENCE MICROSCOPY 10
2.1.1. THE DIFFRACTION LIMIT - RESOLUTION 10
2.1.2. MICROSCOPY METHODS 13
2.2. LOCALIZATION-BASED SUPER-RESOLUTION MICROSCOPY 15
2.2.1. PROBES FOR LOCALIZATION-BASED SUPER-RESOLUTION 16
2.3. MEASURING DYNAMICS IN BIOLOGICAL SAMPLES 22
2.3.1. DIFFUSION, BROWNIAN MOTION AND MEAN SQUARE DISPLACEMENT 22
2.3.2. FLUORESCENCE RECOVERY AFTER PHOTOBLEACHING - FRAP 25
2.3.3. FLUORESCENCE CORRELATION SPECTROSCOPY - FCS 26
2.3.4. SINGLE PARTICLE TRACKING -SPT 28
2.3.5. SUPER-RESOLUTION TRACKING 29
2.3.6. CHOLESTEROL DEPLETION 30
3. MATERIALS AND METHODS 32
3.1. LABELING PROTEINS OF INTEREST 32
3.1.1. IMMUNOSTAINING 32
3.1.2. DNA 32
3.1.3. GENETIC TAGS 33
HTTP://D-NB.INFO/1031575995
IMAGE 2
3.2. ANALYZING SUPER-RESOLUTION DATA 34
3.2.1. DRIFT CONTROL 34
3.2.2. ANALYZING TRACKING DATA 35
4. PUBLICATIONS 37
5. DISCUSSION 38
5.1. SUPER-RESOLUTION FLUORESCENCE MICROSCOPY FOR THE ANALYSIS O F
BIOLOGICAL TARGETS 38
5.1.1. CHOOSING A PROBE 38
5.1.2. PALM, C/STORM OR QDTI? 40
5.1.3. TRACKING PROTEIN DYNAMICS 42
5.1.4. THE OBSERVER S INFLUENCE 44
5.1.5. PROBE SIZE IN MEMBRANE TRACKING AND THE INFLUENCE O F
MICRODOMAINS 44
5.1.6. TRACKING METHODS FOR MEMBRANE COMPONENTS 45
5.2. APPLIED PROJECTS - INTEGRATIVE SUMMARY O F THE RESULTS 47
5.2.1. TNF RECEPTOR QUANTIFICATION 47
5.2.2. LIVE-CELL C/STORM 5 1
5.2.3. TRACKING TNF-R1 WITH SPTPALM: THE ROLE O F LIPID RAFTS 54
6. OUTLOOK 6 1
6.1. MULTICOLOR IN SUPER-RESOLUTION 6 1
6.1.1. TEMPORAL AND SPATIAL COLOCALIZATION 6 1
6.2. LABELING STRATEGIES FOR THE QUANTIFICATION O F MOLECULES 62
6.3. TRACKING CELLULAR DYNAMICS 62
6.4. TRACKING NF-KB AND TRANSCRIPTION DYNAMICS 63
7. PROTOCOLS 65
7.1. VECTOR DESIGN AND DNA AMPLIFICATION 65
7.1.1. VECTORS 65
7.1.2. PRIMERS 66
7.1.3. PCR PROGRAM AND SAMPLE CONDITIONS 66
7.1.4. AGAROSE GEL 66
IMAGE 3
7.1.5. DNA DIGEST 66
7.1.6. E. COLI CULTURE 66
7.2. MAMMALIAN CELL CULTURE 68
7.2.1. HELA CULTURE 68
7.3. CELLULAR LABELING 68
7.3.1. EDHFR LABELING 68
7.3.2. QUANTUM DOT IMMUNOSTAIN 68
7.3.3. CELL TRANSFECTION 69
7.3.4. LUCIFERASE/RENILLA MEASUREMENT 69
7.4. MICROSCOPY 69
7.4.1. CFSTORM SWITCHING BUFFER 69
7.4.2. CELL PREPARATION 70
7.4.3. CONFOCAL LASER SCANNING MICROSCOPY - Q.DTI 70
7.4.4. DSTORM AND PALM 7 1
8. ABBREVIATIONS 72
9. LITERATURE 74
10. ACKNOWLEDGEMENTS 86
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| illustrated | Illustrated |
| indexdate | 2024-07-10T00:25:17Z |
| institution | BVB |
| language | English |
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| physical | 86 S. Ill., graph. Darst. |
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| publishDate | 2012 |
| publishDateSearch | 2012 |
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| spelling | Heidbreder, Meike Verfasser (DE-588)1027212484 aut Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells von Meike Heidbreder Assoziierung und Aktivierung des TNF-Rezeptor I untersucht mit Einzelmolekül-Tracking und hochauflösender Mikroskopie in lebenden Zellen 2012 86 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Würzburg, Univ., Diss., 2012 Fluoreszenzmikroskopie (DE-588)4290958-2 gnd rswk-swf Tumor-Nekrose-Faktor alpha (DE-588)4323543-8 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Tumor-Nekrose-Faktor alpha (DE-588)4323543-8 s Fluoreszenzmikroskopie (DE-588)4290958-2 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-73191 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-73191 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025352071&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Heidbreder, Meike Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells Fluoreszenzmikroskopie (DE-588)4290958-2 gnd Tumor-Nekrose-Faktor alpha (DE-588)4323543-8 gnd |
| subject_GND | (DE-588)4290958-2 (DE-588)4323543-8 (DE-588)4113937-9 |
| title | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells |
| title_alt | Assoziierung und Aktivierung des TNF-Rezeptor I untersucht mit Einzelmolekül-Tracking und hochauflösender Mikroskopie in lebenden Zellen |
| title_auth | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells |
| title_exact_search | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells |
| title_full | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells von Meike Heidbreder |
| title_fullStr | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells von Meike Heidbreder |
| title_full_unstemmed | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells von Meike Heidbreder |
| title_short | Association and Activation of TNF-Receptor I Investigated with Single-Molecule Tracking and Super-Resolution Microscopy in Live Cells |
| title_sort | association and activation of tnf receptor i investigated with single molecule tracking and super resolution microscopy in live cells |
| topic | Fluoreszenzmikroskopie (DE-588)4290958-2 gnd Tumor-Nekrose-Faktor alpha (DE-588)4323543-8 gnd |
| topic_facet | Fluoreszenzmikroskopie Tumor-Nekrose-Faktor alpha Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-73191 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025352071&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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