Abstract
Open-source Multiwavelength Analytical Ultracentrifugation (MWL-AUC) detection systems have been evolving for over a decade. Continual advances emerging out of several research groups have brought the instrumentation technology to increasingly higher levels of performance. The capabilities of MWL-AUC have been documented in many publications, demonstrating the applicability of broad spectrum absorbance acquisitions in analytical ultracentrifugation to a wide array of scientific fields. Despite numerous examples of the usefulness and unique advantages of MWL-AUC, the adoption of the technology by more research groups has been slow. The complexity of the hardware, integration within an ultracentrifuge platform and lack of practical construction and operational information is the likely source of reluctance. Here, we clearly describe the challenges facing a researcher considering adopting MWL-AUC technology in their own laboratories, and provide the information necessary to implement and operate a MWL-AUC system. The discussion includes details of detector assembly, optical alignment, and acquisition parameter settings necessary to achieve high quality experimental results.
Highlights
Past publications on Multiwavelength Analytical Ultracentrifugation (MWL-AUC) have described the overarching principles, demonstrated the technical achievements and illustrated the potential uses of this technology
Once the arm is in place, the lower illumination assembly is fastened to the sliding stage sliding rail assembly at 101.6 mm from the center of the second imaging off-axis parabolic mirror (OAPM) to plane of the CCD
The cross-bars and and cross-hairs and the first imaging OAPM is installed in the focusing assembly (Figure 4d #37), including the 90° flat mirror (Figure 4d #38) and the lower iris
Summary
Past publications on MWL-AUC have described the overarching principles, demonstrated the technical achievements and illustrated the potential uses of this technology. We hope to present a realistic portrayal of the challenges a researcher faces when considering implementing one of these systems in their own laboratory, in addition to many of the critical aspects of the installation that we have found necessary for successful instrument operation demonstrated in past publications [2,6,7,8]. This includes a description of the necessary tools and expertise to build and install the detector, followed by a protocol for alignment and calibration.
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