Development of MWL-AUC/CCD-C-AUC/SLS-AUC detectors for the analytical ultracentrifuge [Elektronische Ressource] / von Engin Karabudak

Engin Karabudak

Informations

• Publié par: universitat_potsdam
• Publié le: 01 janvier 2009
• Nombre de lectures: 33
• Poids de l'ouvrage: 7 Mo

Extrait

Max-Planck Institut für Kolloid und Grenzflächenforschung








Development of

MWL-AUC / CCD-C-AUC / SLS-AUC
Detectors

for the Analytical Ultracentrifuge








Dissertation
zur Erlangung des akademischen Grades
„doctor rerum naturalium“
(Dr. rer. nat.)
in der Wissenschaftsdisziplin „Kolloidchemie“



eingereicht an der
Mathematisch-Naturwissenschaftlichen Fakultät
Universität Potsdam



von
Engin Karabudak


Potsdam, im Mai 2009 This work is licensed under a Creative Commons License:
Attribution - Noncommercial - Share Alike 3.0 Germany
To view a copy of this license visit
http://creativecommons.org/licenses/by-nc-sa/3.0/de/deed.en










































Published online at the
Institutional Repository of the University of Potsdam:
URL http://opus.kobv.de/ubp/volltexte/2009/3992/
URN urn:nbn:de:kobv:517-opus-39921
http://nbn-resolving.org/urn:nbn:de:kobv:517-opus-39921 TABLE OF CONTENTS:

CHAPTER 1 : THESIS INTRODUCTION........................................................................................................ 1
CHAPTER 2 : GENERAL REVIEW OF AUC ................................................................................................. 6
2.1. GENERAL ANALYTICAL ULTRACENTRIFUGATION................................................................................. 6
2.2. INSTRUMENTATION FOR COMMERCIAL AUC ........................................................................................ 8
2.2.1. Mechanical Parts ............................................................................................................................ 8
2.2.1.1. Rotors ......................................................................................................................................................... 8
2.2.1.2. Cells............................................................................................................................................................ 9
2.2.2. Electronic Parts ............................................................................................................................ 11
2.2.3. Optical Parts ................................................................................................................................. 12
2.2.3.1. Absorbance Optics.................................................................................................................................... 13
2.2.3.2. Interference Optics ................................................................................................................................... 14
2.3. THEORY OF ANALYTICAL ULTRACENTRIFUGE.................................................................................... 16
2.4. OTHER DETECTORS FOR XL-AUC...................................................................................................... 19
2.4.1. Schlieren Optics............. 20
2.4.2. Fluorescence Detector .................................................................................................................. 21
2.4.3. Turbidity Detector......................................................................................................................... 21
2.5. TYPES OF EXPERIMENTS WITH AUC................................................................................................... 22
2.5.1. Sedimentation Velocity Experiment............................................................................................... 22
2.5.2. Sedimentation Equilibrium Experiment ........................................................................................ 23
2.5.3. Band (Zone) Centrifugation .......................................................................................................... 24
CHAPTER 3 : STATIC LIGHT SCATTERING DETECTOR FOR ANALYTICAL
ULTRACENTRIFUGATION (SLS-AUC) ....................................................................................................... 26
3.1. INTRODUCTION ................................................................................................................................... 26
3.2. EXPERIMENTAL TESTS........................................................................................................................ 27
3.2.1. LAAPD with home-made power supply......................................................................................... 27
3.2.2. Modifying the commercial power supply of the LAAPD ............................................................... 29
3.2.3. First Prototype of SLS-AUC..... 29
3.2.4. Tests with the prototype SLS-AUC ................................................................................................ 30
CHAPTER 4 : CCD CAMERA DETECTOR FOR THE ANALYTICAL ULTRACENTRIFUGE (CCD-
C-AUC) ................................................................................................................................................................ 31
4.1. INTRODUCTION ................................................................................................................................... 31
4.2. EXPERIMENTAL TESTS........................................................................................................................ 32
4.2.1. Resolution Test.............................................................................................................................. 33
4.2.2. Monochromator of the Optima XL-I.............................................................................................. 34
4.2.3. Illumination test with SK2048DDE inside AUC with Xenon flash lamp....................................... 36
4.2.4. Owl Camera inside AUC with Xenon flash lamp .......................................................................... 36
4.2.5. Tests with Constant Light Sources in the Optima XL-I ................................................................. 36
4.2.6. Illumination tests on the Optical Bench with 75 W light............................................................... 37
4.2.7. Construction of prototype test setup.............................................................................................. 37
4.2.8. Constant Light Source from Aviv Biomedical ............................................................................... 39
4.2.9. ant Light Source, Test Setup with Monochromator.............................................................. 40
4.2.10. First Prototype CCD-C-AUC, taking UV/Vis spectra 41
CHAPTER 5 : MULTIWAVELENGTH DETECTOR FOR ANALYTICAL ULTRACENTRIFUGE
(MWL-AUC) ....................................................................................................................................................... 45
5.1. INTRODUCTION ................................................................................................................................... 45
5.2. IMPROVEMENT OF THE MULTIWAVELENGTH DETECTOR...................................................................... 46
5.2.1. Flash Lamp.................................................................................................................................... 46
5.2.2. Detector Arm and Spectrometer Mount ........................................................................................ 47
5.2.3. Imaging Optics.............................................................................................................................. 48
5.2.4. Optical Tests.................................................................................................................................. 48
5.3. RESULTS.............. 49
5.3.1. General Aspects ............................................................................................................................ 49
5.3.2. Radial Resolution .............................................................................................................. 52
5.3.3. Wavelength Accuracy.................................................................................................................... 53
ii5.3.4. Absorbance Accuracy and Linearity ............................................................................................. 55
5.3.5. Intrinsic Noise of the Data ............................................................................................................ 57
5.4. DISCUSSION ........................................................................................................................................ 59
CHAPTER 6 : BIOLOGICAL APPLICATION OF MWL-AUC: PROTEIN MIXTURE.......................... 60
6.1. INTRODUCTION.... 60
6.2. RESULTS AND DISCUSSION ................................................................................................................. 60
CHAPTER 7 : INDUSTRIAL APPLICATION OF MWL-AUC: INVESTIGATION OF Β-CAROTENE-
GELATIN COMPOSITE PARTICLES....... 67
7.1. INTRODUCTION.... 67
7.2. MATERIAL AND METHODS.................................................................................................................. 68