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Documents
2008
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153
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English
Ebook
2008
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Publié le
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Poids de l'ouvrage
16 Mo
Publié par
Publié le
01 janvier 2008
Nombre de lectures
12
Langue
English
Poids de l'ouvrage
16 Mo
Fluorescent Multiple Chemical Sensing using Time-
Domain Fluorescence Lifetime Imaging
Dissertation zur Erlangung des Doktorgrads der Naturwissenschaften
(Dr. rer. nat.)
der Fakultät Chemie und Pharmazie
der Universität Regensburg
vorgelegt von
Stefan Nagl
Regensburg, im Mai 2008
Diese Doktorarbeit entstand in der Zeit vom Juni 2004 bis zum April 2008 am Institut für
Analytische Chemie, Chemo- und Biosensorik der Universität Regensburg.
Die Arbeit wurde angeleitet von Prof. Otto S. Wolfbeis
Promotionsgesuch eingereicht am: 27.05.2008
Kolloquiumstermin: 19.06.2008
Prüfungsausschuß: Vorsitzender: Prof. Hans-Helmut Kohler
Erstgutachter: Prof. Otto S. Wolfbeis
Zweitgutachter: Prof. Achim Göpferich
Drittprüfer: Prof. Bernhard Dick
Contents
CONTENTS
CHAPTER 1
Introduction
1.1. Time-resolved Fluorescence Imaging...........................................................................1
1.2. Chemical Sensors and Biosensors.................................................................................4
1.3. Optical Multiple Chemical Sensing ............................................................................10
1.4. Sensor Miniaturization and Microarray Technology ...............................................14
1.5. Aim of the Research......................................................................................................16
1.5. References ......................................................................................................................18
CHAPTER 2
Record Response Optical Trace Oxygen Sensing and Imaging
2.1. Introduction...................................................................................................................22
2.2. Results and Discussion..................................................................................................23
2.2.1. Fluorescence spectra...........................................................................................................23
2.2.2. Fluorescence and phosphorescence spectra at low temperatures.......................................25
2.2.3. Singlet oxygen luminescence spectra and lifetimes...........................................................26
2.2.4. Scanning electron micrographs..........................................................................................29
2.2.5. Fluorescence lifetime imaging...........................................................................................30
2.2.6. Comparison of oxygen quenching efficiencies..................................................................33
2.3. Conclusion ....................................................................................................................34
2.4. Experimental Section....................................................................................................35
2.5. References......................................................................................................................36
I Contents
CHAPTER 3
A Dual Fluorescence Sensor for Trace Oxygen and Temperature with a
Large Temperature Range and Unmatched Oxygen Sensitivity
3.1. Introduction...............................................................................................................39
3.2. Experimental Section ................................................................................................42
3.2.1. Materials.............................................................................................................................42
3.2.2. Methods..............................................................................................................................42
3.3. Results and Discussion ..............................................................................................45
3.4.1. Composition of the dual sensors ...................................................................................45
3.4.2. Luminescence spectra...................................................................................................47
3.4.3. Calibration of the dual sensors .....................................................................................49
3.4.4. Response times of the dual sensors to oxygen..............................................................56
3.4.5. Derivation and validation of a bivariate calibration function for all temperatures and
oxygen concentrations covered.....................................................................................58
3.4. Conclusion......................................................................................................................63
3.5. References.......................................................................................................................64
CHAPTER 4
A Method for Simultaneous Luminescence Sensing and Imaging of Two
Species Using Optical Probes of Different Luminescence Decay Time
4.1. Introdcution...............................................................................................................70
4.2. Results and Discussion ..............................................................................................72
4.2.1. Experimental design ..........................................................................................................72
4.2.2. Material selection...............................................................................................................75
4.2.3. Calibration of the temperature sensitivity ..........................................................................77
4.2.4. Calibration of the oxygen sensitivity .................................................................................79
II Contents
4.2.5. Measurement of oxygen consumption caused by enzymatic catalysis at varying
temperatures.......................................................................................................................82
4.3. Conclusion...................................................................................................................... 83
4.4. Experimental Section ................................................................................................84
4.4.1. Materials.............................................................................................................................84
4.4.2. Preparation of the dual sensing film...................................................................................85
4.4.3. Calibration of the dual sensor ............................................................................................86
4.4.4. Enzymatic oxygen consumption measurements ................................................................87
4.5. References......................................................................................................................89
CHAPTER 5
Luminescent Polymer Nanoparticles as Probes for Protein, Oxygen and
Temperature
5.1. Introduction...............................................................................................................92
5.2. Experimental part .....................................................................................................94
5.2.1. Materials.............................................................................................................................94
5.2.2. Polymer syntheses..............................................................................................................95
5.2.3. Nanoparticle syntheses.......................................................................................................96
5.2.4. Instruments.........................................................................................................................98
5.3. Metalloporphyrin-doped phosphorescent PD nanoparticles as optical probes.....100
5.3.1. Platinum porphyrin-doped nanospheres displaying FRET to red-emitting cyanine
dyes......................................................................................................................... .........100
5.3.2. Palladium benzoporphyrin-doped nanospheres for NIR applications..............................106
5.4. Temperature-sensitive doped PMAN nanospheres................................................108
5.4.1. Optical spectra and nanoparticle size...............................................................................109
5.4.2. Temperature sensitivity....................................................................................................109
5.5. Dye-doped polystyrene-based nanobeads for oxygen sensing........