Abstract
The advantages of simultaneously detecting multiple wavelengths in ultracentrifugation experiments are obvious, especially for interacting systems. In addition, the detection of the wavelength dependence of turbidity opens up the possibility to obtain independent information on the particle size in addition to the usual sedimentation coefficient distribution for colloidal systems. We therefore made an effort to develop a fast UV/Vis detector, which is able to simultaneously detect the range from 200–800 nm. This is possible by the use of a modern CCD chip based generation of UV-Vis spectrometers, which translates the dispersed white light onto a CCD chip, where each pixel corresponds to a particular wavelength. In addition to the simultaneous detection of a large number of wavelengths in the range 200–800 nm, also with non integer values, these spectrometers are very fast. Current typical spectrum scan times with the necessary scan quality in the ultracentrifuge are in the range of 100 ms but this time can be significantly shortened down to 3 ms for higher light intensities and even down to 10 μs for a new generation of CCD chip based spectrometers.
The introduction of a fiber based UV-Vis optics into a preparative XL-80K ultracentrifuge with the associated hardware developments will be described as a first generation prototype. In this study, we use a wavelength dependent optical lens system instead of the necessary but more complex wavelength independent mirror optical system for a first check on possibilities and limitations of the optical system. First examples for biopolymers and latexes will be presented and compared to those obtained in the commercial XL-A ultracentrifuge. Already the fast detection enables completely new possibilities like the determination of a particle size distribution in a few minutes. Multiwavelength detection at constant position in dependence of time will be demonstrated, which is an important mode for the use of speed profiles for very polydisperse samples. Also, the use of radial multiwavelength scans will be demonstrated producing a three dimensional data space for monitoring the sedimentation via radial scans with multiwavelength detection. However, despite the advantages, the current problems with the detector will also be discussed including the main problem that much intensity is lost in the important UV range as a result of fiber coupling and bending.
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References
Svedberg T, Pedersen KO(1940) The Ultracentrifuge. Clarendon Press, Oxford
Lloyd PH (1974) Optical methods in Ultracentrifugation, Electrophoresis, and Diffusion: with a guide to the interpretation of records. Clarendon Press, Oxford
Schmidt B, Riesner D (1992) A Fluorescence Detection System for the Analytical Ultracentrifuge and its Application to Proteins, Nucleic Acids, Viroids and Viruses. In: Harding SE et al. (ed) Analytical Ultracentrifugation in Biochemistry and Polymer Science. Royal Society of Chemistry, Cambridge, p 176
MacGregor IK, Laue TM (1999) Biophys J 76:357
MacGregor IK, Anderson AL, Laue TM (2004) Biophys Chem 108:165
Cantow HJ (1964) Makromol Chem 70:130
Scholtan W, Lange H (1972) Kolloid Z Z Polym 250:782
Müller HG (1989) Colloid Polym Sci 267:1113
Mächtle W (1992) Analysis of Polymer Dispersions with an Eight-Cell-AUC-Multiplexer: High Resolution Particle Size Distribution and Density Gradient Techniques. In: Harding SE et al. (ed) Analytical Ultracentrifugation in Biochemistry and Polymer Science. Royal Societyof Chemistry, Cambridge, p 147
Klodwig U, Mächtle W (1989)Colloid Polym Sci 267:1117
Clewelow AC, Errington N, Rowe AJ (1997) Eur Biophys J 25:311
Holtus G, Borchard W (1989) Colloid and Polymer Science 267:1133
Cölfen H, Borchard W (1994) Progr Colloid Polym Sci 94:90
Kisters D, Borchard W (1999) Progr Colloid Polym Sci 113:10
Laue TM (1992) On-Line data aquisition and analysis from the Rayleigh interferometer. In: Harding SE et al. (ed) Analytical Ultracentrifugation in Biochemistry and Polymer Science. Royal Society of Chemistry, Cambridge, p 63
Börger L, Lechner MD, Stadler M (2004) Progr Colloid Polym Sci 127:19
Mächtle W (1999) Progr Colloid Polym Sci 113:1
Flossdorf J, Schillig H, Schindler KP (1978) Makromol Chem 179:1617
Giebeler R (1992) The Optima XL-A: A New Analytical Ultracentrifuge with a Novel Precision Absorption Optical System. In: Harding SE et al. (ed) Analytical Ultracentrifugation in Biochemistry and Polymer Science. Royal Society of Chemistry, Cambridge, p 16
Flossdorf J (1980) Makromol Chem 181:715
Cölfen H, Borchard W (1994) Ultrasensitive Schlieren Optical System. In: Bonner RF et al. (ed) Progress in Biomedical Optics, Vol 2136. SPIE Bellingham, Washington, p 307
Schuck P (2000) Biophys J 78:1606
Stafford WF (1992) Anal Biochem 203:295
Johnson ML, Correia JJ, Yphantis DA, Halvorson (1981) Biophys J 36:575
McRorie DK, Voelker PJ (1993) Self-Associating Systems in the Analytical Ultracentrifuge. Beckman Instruments California ( http://www.beckman.com/Literature/BioResearch/362784.pdf )
Demeler B, van Holde KE (2004) Anal Biochem 335:279 and Ultrascan Website: http://www.ultrascan.uthscsa.edu/
Mächtle W (1991) Progr Colloid Polym Sci 86:111
Voelker P (1995) Progr Colloid Polym Sci 99:162
Cölfen H, Pauck T, Antonietti M (1997) Progr Colloid Polym Sci 107:136
Schuck P (2003) Anal Biochem 320:104
Cölfen H, Völkel A (2003) Eur Biophys J 32:432
Global analysis is also possible with the SEDPHAT software by Peter Schuck. See the following Website for further details: http://www.analyticalultracentrifugation.com/sedphat/sedphat.htm
Vistica J, Dam J, Balbo A, Yikilmaz E, Mariuzza RA, Rouault TA, Schuck P (2004) Anal Biochem 326:234
Balbo A, Minor KH, Velikovsky CA, Mariuzza RA, Peterson CB, Schuck P (2005) Proc Natl Acad Sci USA 102 1:81
Gledhill RJ (1962) J Phys Chem 66:458
Bateman JB, Weneck EJ, Eshler DC (1959) J Colloid Sci 14:308
Heller W, Bhatnagar HL, Nakagaki M (1962) J Chem Phys 36:1163
Hosono M, Sugii S, Kusudo O, Tsuji W (1973) Bull Inst Chem Res, Kyoto Univ 51:104
http://www.oceanoptics.com/technical/engineering/USB2000%20OEM%20Data%20Sheet.pdf
http://www.oceanoptics.com/technical/engineering/OEM%20Data%20Sheet%20–%20HR4000.pdf
www.lot-oriel.com/ccd
Cölfen H (1994) Bestimmung thermodynamischer und elastischer Eigenschaften von Gelen mit Hilfe von Sedimentations-gleichgewichten in einer Analytischen Ultrazentrifuge am Beispiel des Systems Gelatine/Wasser. Verlag Köster, Berlin
Visit the following website: http://www.usa.hamamatsu.com/en/products/electron-tube-division/light-sources/xenon-flash-lamps.php
Cölfen H, Borchard W (1994) Anal Biochem 219:321
Laue TM (1981) PhD Dissertation, Univ Of Connecticut, Storrs, CT,USA
Rockholt DL, Royce CR, Richards (1976) Biophys Chem 5:55
Laue TM, Domanic RA, Yphantis DA (1983) Anal Biochem 131:220
Yphantis DA, Laue TM, Anderson IA (1983) Anal Biochem 143:95
Mächtle W, Klodwig U (1979) Makromol Chem 180:2507
Mächtle W, Klodwig U (1976) Makromol Chem 177:1607
Sedlack U, Lechner MD (1995) Progr Colloid Polym Sci 99:136
Ortlepp B, Panke D (1991) Progr Colloid Polym Sci 86:57
Schindler KP (1980) Avail NTIS. Report (Order No. PB81-167140):157
Flossdorf J, Schillig H, Schindler KP (1980) J Phys E: Scientific Instruments 13:647
Kuhnert R, Boedel E, Stegemann H, Wastl G (1973) CZ-Chem Tech 2:441
Müller HG (2004) Progr Colloid Polym Sci 127:9
Stafford WE, Braswell EH (2004) Biophys Chem 108:273
Müller HG (1989) Colloid Polym Sci 267:1113
Mächtle W (1999) Biophys J 76:1080
Mächtle W (1988) Angew Makromol Chem 162:35
Acknowledgments
We thank the BASF AG, Ludwigshafen and the Max-Planck-Society for financial support of this work. We also thank Dr. Walter Mächtle, BASF, for useful discussions and Dr. Borries Demeler, UTHCSA, San Antonio for writing a data visualization program to display the multiwavelength data. The mechanical workshop of the BASF is acknowledged for the modification of the XL heatsink and for building the basic detector arm. Andreas Kretzschmar, mechanical workshop MPI-KGF, is acknowledged for multiple modifications of the detector arm. Henryk Pitas, electrical workshop MPI-KGF, is thanked for help with electrical problems. Dr. K. Tauer (MPI-KGF) is acknowledged for the latex samples and Dr. Neil Robinson (UTHCSA) for the cytochrome samples. Finally, we thank Antje Völkel for helpful assistance and Prof. Dr. Dr. h.c. Markus Antonietti for the overall support of this project.
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Bhattacharyya, S.K. et al. Development of a Fast Fiber Based UV-Vis Multiwavelength Detector for an Ultracentrifuge. In: Wandrey, C., Cölfen, H. (eds) Analytical Ultracentrifugation VIII. Progress in Colloid and Polymer Science, vol 131. Springer, Berlin, Heidelberg. https://doi.org/10.1007/2882_002
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DOI: https://doi.org/10.1007/2882_002
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