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Optimized Imaging of Protein Gels Stained with Coomassie,,,Brilliant Blue Dye

filter for the Eagle Eye II still video system is the Evergreen Filter, an optical bandpass filter designed for SYBR Green nucleic acid stain and FLASH CAT applications.1 The Evergreen Filter maximally overlaps the emission spectra of both of these green fluorophores, assuring optimal sensitivity and signal capture efficiency.

In this report, we describe an optical filter specifically designed for use with Coomassie brilliant blue-stained protein gels. Unlike ethidium bromide, SYBR Green and BODIPY stains, Coomassie brilliant blue is not a fluorescent dye. Proteins stained with Coomassie blue appear blue in color because the stain absorbs non-blue wavelengths of visible light while reflecting blue light. With reflected light, contrast control filters can be used to increase the contrast between an object and its background.2 In this way, the apparent brightness of a colored object can be increased by photographing it through a filter of a complementary color. We have designed a contrast control filter with a narrow bandpass for visualizing Coomassie blue-stained protein gels. As are the other Eagle Eye optical filters, the Coomassie blue bandpass filter is coated to ensure high image quality.

Comparison of Optical Filters

In order to compare the performance of optical filters when imaging Coomassie blue-stained protein gels, various protein preparations were fractionated through a 4-20% SDS-PAGE mini-gel and stained with Coomassie brilliant blue dye using standard conditions.3 The gel was positioned on top of a white-light box and imaged using white-light transillumination. The Eagle Eye II still video system was fitted with either the ethidium bromide optical filter or the Coomassie blue bandpass filter. Images were also taken using each filter with overhead white-light illumination while the gel was resting on top of a pie
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