Microscopy & Microtechniques
Using A CCD-Based Imager To Scan Pace Gels Provides A Quick And Sensitive Method Of Analysis
Mar 23 2007
Author: Florence Goubet, PhD, Paul Dupree, PhD, Alexandra Godlee and Paru Oatey, DPhil on behalf of Unassigned Independent Article
Analysis of polysaccharides can be problematic because of the need for complex methods and/or expensive equipment such as chromatography and mass spectrometry [1,2], NMR [3], capillary electrophoresis [4], and Fourier transform infrared spectroscopy [3,5]. These techniques are ideal for a particular polysaccharide or application, but they have limitations, such as the need for pure compounds or relatively large sample quantity. Recently, a method known as PACE (polysaccharide analysis using carbohydrate gel electrophoresis) has been described [6,7]. This can be used in a high throughput manner and is applicable to small quantities of material. PACE relies on derivatising reducing ends of sugars and oligosaccharides with a fluorophore label, followed by polyacrylamide gel electrophoresis. To obtain sensitive detection of fluorescent signals on PACE gels generally requires laser based scanners which can be expensive; limited to fluorescence applications; or require large amounts of bench space. To overcome these difficulties, this article describes how CCD-based technology can be used to detect the fluorescent signal produced on a PACE gel and the sensitivity this type of system can achieve.
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