Microscopy & Microtechniques
New Application of Image Analyser Offers Rapid, Accurate Detection of Nanogram Amounts of Protein
Nov 06 2006
Syngene's technical team used a Dyversity system with a Cy dye lighting module, a dual wavelength transilluminator, a blue light converter, UV, long pass, Cy3 dye and SG emission filters to image 1D acrylamide gels stained with Sypro Ruby, containing 1000-0.1ng of molecular weight standard PeppermintStick? (Invitrogen). The gels were imaged under four conditions: Cy2 excitation with a Cy3 emission filter; Cy2 excitation with a UV emission filter; medium-wave UV excitation with a UV filter and medium-wave UV excitation with a blue light converter and SG filter.
The results showed using the medium-wave UV excitation with a blue light converter and SG filter, Dyversity could detect 5ng of Sypro Ruby stained protein in 4 seconds. The Cy2 illumination with either Cy3 or long pass emission filter allowed detection of 0.5ng of protein in two seconds, but the best results were obtained using the medium-wave UV and a UV filter which detected 0.5ng of protein, in under one second, unrivalled imaging performance for a CDD-based analyser.
Dyversity can acquire images of small amounts of fluorescently labelled protein so rapidly because it has a 90 micron resolution, 16-bit CCD camera. This provides Dyversity with the fastest capture time per channel for Cy dyes of any CCD system on the market today, and means it is an excellent alternative to a laser-based scanner.
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