Abstract
A new concept for multiwavelength detection in analytical ultracentrifugation is demonstrated, based on the technology of an Andor iStar intensified CCD camera and constant illumination source. The camera is coupled to an Andor Shamrock spectrograph providing a spectrometer system capable of UV/vis acquisitions with wavelengths from 190 to 790 nm. The details of the camera functions are described, and the essential operational modes demonstrated with proof of principle measurements. The Andor spectrometer system is proven to be much more sensitive than previous AUC detectors. In fact, it is so sensitive in high gain modes that signal quality is limited by photon shot-noise. Acquisition strategies are presented for detection systems working with low-light illumination sources, and the necessity of overcoming shot-noise limited signal quality is revealed. Several illumination optical concepts are tested and it is shown that when sufficient illumination is applied the detection system is capable of outperforming commercial and academic detectors previously reported in terms of signal-to-noise. A path forward for overcoming the remaining challenges is provided. The findings in this manuscript will be applicable to any type of high sensitivity detector design.
Highlights
Recent years have brought new developments in ultracentrifuge technology, such as the introduction of the Centrifugal Fluid Analyzer (CFA) from Spin Analytical, opening up new opportunities for analytical ultracentrifugation (AUC) detector development
The Open AUC MWL absorbance detectors previously described have been designed to work within the BeckmanCoulter preparative style centrifuges.[2,3,4,5,6]
The spectrometer system used here is composed of an iStar Intensi ed Charge Coupled Device (ICCD) 2D array camera (DH320T-25F-04) with Gen 2 Intensi er and Shamrock 193i spectrograph from Andor Technology Ltd., where the camera's horizontal pixel array captures the spectral domain, and the vertical pixel array images the spatial dimension along the length of the spectrograph entrance slit
Summary
Recent years have brought new developments in ultracentrifuge technology, such as the introduction of the Centrifugal Fluid Analyzer (CFA) from Spin Analytical, opening up new opportunities for analytical ultracentrifugation (AUC) detector development. The Open AUC MWL absorbance detectors previously described have been designed to work within the BeckmanCoulter preparative style centrifuges.[2,3,4,5,6] The detectors are mounted in the centrifuge vacuum chamber and light is introduced via optical bers. This imposes hardware constraints, University of Konstanz, Physical Chemistry, Department of Chemistry, Universitatsstraße 10, Konstanz 78457, Germany. The illumination of a D–W assembly is signi cantly lower than xenon or xenon–mercury arc lamps, but is able to provide balanced spectral output from the UV through the visible
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