Optimizing microscopic Förster resonance energy transfer measurements:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Jena
[2019]
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | 134 Blätter Illustrationen 29 cm |
Internformat
MARC
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| 100 | 1 | |a Langenstück, Teresa |d 1986- |e Verfasser |0 (DE-588)120097414X |4 aut | |
| 245 | 1 | 0 | |a Optimizing microscopic Förster resonance energy transfer measurements |c von Teresa Langenstück |
| 264 | 1 | |a Jena |c [2019] | |
| 300 | |a 134 Blätter |b Illustrationen |c 29 cm | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |b Dissertation |c Friedrich-Schiller-Universität Jena |d 2019 | ||
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Datensatz im Suchindex
| _version_ | 1804180949120843776 |
|---|---|
| adam_text | LIST
OF
CONTENTS
1.
I
NTRODUCTION
................................................
1
1.1.
PROTEIN-PROTEIN
INTERACTION
RESEARCH
...........................................................................
1
1.2.
PRINCIPLES
OF
FLUORESCENCE
.............................................................................................
3
1.2.1.
INTRODUCTION
TO
FLUORESCENCE
............................................................................
3
1.2.2.
FLUORESCENCE
LIFETIME
........................................................................................
4
1.3.
FORSTER
RESONANCE
ENERGY
TRANSFER
(FRET)
................................................................
5
1.4.
MEASUREMENT
OF
FRET
.................................................................................................
11
1.5.
TRIPLE
FRET..................................................................................................................
16
1.6.
FLUORESCENT
PROTEINS
FOR
FRET
....................................................................................
17
2.
A
IMS
OF
THIS
THESIS
.............................................
22
3.
M
ATERIAL
AND
M
ETHODS
.........................................
23
3.1.
DNA
VECTOR
PREPARATION,
TRANSFECTION,
TRANSFORMATION,
AND
CELL
CULTURE
...........................................................................................................................
23
3.1.1.
CONSTRUCTION
OF
EXPRESSION
VECTORS
...............................................................
23
3.1.2.
CALCIUM
PHOSPHATE
MEDIATED
TRANSIENT
TRANSFECTION
OF
EUKARYOTIC
CELLS
...............................................................................................
26
3.1.3.
BACTERIAL
TRANSFORMATION
USING
HEAT
SHOCK
AND
COMPETENT
CELLS
...................................................................................................................
28
3.2.
ISOLATION
OF
FLUORESCENT
PROTEINS
................................................................................
28
3.3.
SPECTROSCOPIC
METHODS
................................................................................................
31
3.3.1.
MEASUREMENT
OF
ABSORPTION
SPECTRA
..............................................................
31
3.3.2.
MEASUREMENT
OF
FLUORESCENCE
EXCITATION
AND
EMISSION
SPECTRA
.............................................................................................................
32
3.4.
FLUORESCENCE
MICROSCOPY
.............................................................................................
33
3.4.1.
CONFOCAL
LASER
SCANNING
MICROSCOPY
..............................................................
33
3.4.2.
FLUORESCENCE
LIFETIME
MEASUREMENTS
............................................................
35
3.5.
FRET
MEASUREMENTS
...................................................................................................
50
3.5.1.
INTENSITY-BASED
FRET
MEASUREMENTS
............................................................
50
3.5.2.
FLUORESCENCE
LIFETIME-BASED
FRET
MEASUREMENTS
.......................................59
4.
R
ESULTS
.....................................................
60
4.1.
CHARACTERIZATION
OF
CYAN
FPS
........................................................................................60
4.1.1.
ABSORPTION-
AND
FLUORESCENCE
SPECTRA
OF
THE
FLUORESCENT
PROTEINS
...............................................................................................
60
LIST
OF
CONTENTS
4.1.2.
TIME
AND
SPECTRALLY
RESOLVED
EXAMINATION
OF
FLUORESCENT
PROTEINS
............................................................................................................
63
4.2.
OPTIMIZING
FRET
MEASUREMENTS
.................................................................................
73
4.2.1.
INTENSITY-BASED
FRET
MEASUREMENTS
............................................................
73
4.2.2.
FLUORESCENCE
LIFETIME
MEASUREMENTS
WITH
TCSPC
........................................86
4.2.3.
OPTIMIZING
A
TCSPC
APPROACH
FOR
TRIPLE
FRET
.............................................. 88
5.
D
ISCUSSION
................................................... 97
5.1.
CHARACTERIZATION
OF
CYAN
FPS
TO
FIND
THE
BEST
DONOR
FLUOROPHORE
FOR
FRET
EXPERIMENTS
.......................................................................................................
97
5.1.1.
SPECTRAL
PROPERTIES
.........................................................................................
97
5.1.2.
MULTIEXPONENTIAL
DECAYS
................................................................................
98
5.1.3.
EFFECTS
OF
BLEACHING
ON
SINGLE
FPS
...................................................................99
5.1.4.
EFFECTS
OF
BLEACHING
ON
FUSION
PROTEINS
........................................................
101
5.1.5.
DATA
ANALYSIS
.................................................................................................
105
5.1.6.
CHOICE
OF
THE BEST
DONOR
FLUOROPHORE
.......................................................... 105
5.2.
OPTIMIZING
FRET
MEASUREMENTS
..............................................................................
106
5.2.1.
USE
OF
COMMERCIAL
SOFTWARE
SUITES
..............................................................
107
5.2.2.
ANALYSIS
OF
THE
SETUP
.....................................................................................
108
5.2.3.
ANALYSIS
OF
THE
DATA
.......................................................................................
110
5.3.
OPTIMIZING
A
TCSPC
APPROACH
FOR
TRIPLE
FRET........................................................ 112
5.4.
SUGGESTIONS
FOR
FRET
MEASUREMENTS
.......................................................................
114
6.
R
EFERENCES
..................................................
116
7.
A
PPENDIX
...................................................
123
7.1.
PROPERTIES
OF
SELECTED
FLUORESCENT
PROTEINS
............................................................. 123
7.2.
USED
SYMBOLS
AND
DEFINITIONS
...................................................................................
127
7.3.
LIST
OF
FIGURES
.............................................................................................................
130
7.4.
LIST
OF
TABLES
..............................................................................................................
132
DANKSAGUNG
..................................................................................................................................
133
EHRENWORTLICHE
ERKLARUNG
..........................................................................................................
134
II
|
| any_adam_object | 1 |
| author | Langenstück, Teresa 1986- |
| author_GND | (DE-588)120097414X |
| author_facet | Langenstück, Teresa 1986- |
| author_role | aut |
| author_sort | Langenstück, Teresa 1986- |
| author_variant | t l tl |
| building | Verbundindex |
| bvnumber | BV046410437 |
| ctrlnum | (OCoLC)1138064981 (DE-599)DNB1200361873 |
| discipline | Biologie |
| format | Thesis Book |
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| genre_facet | Hochschulschrift |
| id | DE-604.BV046410437 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T08:43:53Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-031823064 |
| oclc_num | 1138064981 |
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| physical | 134 Blätter Illustrationen 29 cm |
| publishDate | 2019 |
| publishDateSearch | 2019 |
| publishDateSort | 2019 |
| record_format | marc |
| spelling | Langenstück, Teresa 1986- Verfasser (DE-588)120097414X aut Optimizing microscopic Förster resonance energy transfer measurements von Teresa Langenstück Jena [2019] 134 Blätter Illustrationen 29 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Schiller-Universität Jena 2019 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1200361873/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031823064&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Langenstück, Teresa 1986- Optimizing microscopic Förster resonance energy transfer measurements |
| subject_GND | (DE-588)4113937-9 |
| title | Optimizing microscopic Förster resonance energy transfer measurements |
| title_auth | Optimizing microscopic Förster resonance energy transfer measurements |
| title_exact_search | Optimizing microscopic Förster resonance energy transfer measurements |
| title_full | Optimizing microscopic Förster resonance energy transfer measurements von Teresa Langenstück |
| title_fullStr | Optimizing microscopic Förster resonance energy transfer measurements von Teresa Langenstück |
| title_full_unstemmed | Optimizing microscopic Förster resonance energy transfer measurements von Teresa Langenstück |
| title_short | Optimizing microscopic Förster resonance energy transfer measurements |
| title_sort | optimizing microscopic forster resonance energy transfer measurements |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1200361873/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031823064&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT langenstuckteresa optimizingmicroscopicforsterresonanceenergytransfermeasurements |
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