title: Performance validation of an improved Xenon-arc lamp-based CCD camera system for multispectral imaging in proteomics.
abstract: Advances in gel-based nonradioactive protein expression and PTM detection using fluorophores has served as the impetus for developing analytical instrumentation with improved imaging capabilities. We describe a CCD camera-based imaging instrument, equipped with both a high-pressure Xenon arc lamp and a UV transilluminator, which provides broad-band wavelength coverage (380-700 nm and UV). With six-position filter wheels, both excitation and emission wavelengths may be selected, providing optimal measurement and quantitation of virtually any dye and allowing excellent spectral resolution among different fluorophores. While spatial resolution of conventional fixed CCD camera imaging systems is typically inferior to laser scanners, this problem is circumvented with the new instrument by mechanically scanning the CCD camera over the sample and collecting multiple images that are subsequently automatically reconstructed into a complete high-resolution image. By acquiring images in succession, as many as four different fluorophores may be evaluated from a gel. The imaging platform is suitable for analysis of the wide range of dyes and tags commonly encountered in proteomics investigations. The instrument is unique in its capabilities of scanning large areas at high resolution and providing accurate selectable illumination over the UV/visible spectral range, thus maximizing the efficiency of dye multiplexing protocols.
authors.affiliation: PerkinElmer Life and Analytical Sciences, Seer Green, UK.
authors.forename: Elaine
authors.initials: E
authors.lastname: Scrivener
authors.forename: Brett A
authors.initials: BA
authors.lastname: Boghigian
authors.forename: Eva
authors.initials: E
authors.lastname: Golenko
authors.forename: Alla
authors.initials: A
authors.lastname: Bogdanova
authors.forename: Peter
authors.initials: P
authors.lastname: Jackson
authors.forename: Alvydas
authors.initials: A
authors.lastname: Mikulskis
authors.forename: Eric
authors.initials: E
authors.lastname: Denoyer
authors.forename: Patrick
authors.initials: P
authors.lastname: Courtney
authors.forename: Mary F
authors.initials: MF
authors.lastname: Lopez
authors.forename: Wayne F
authors.initials: WF
authors.lastname: Patton
mesh_terms: D029681:Arabidopsis Proteins / Q000032:analysis; D004396:Coloring Agents; D015902:Gamma Cameras; D005782:Gels; D007091:Image Processing, Computer-Assisted; D010750:Phosphoproteins / Q000032:analysis* / Q000648:ultrastructure; D011506:Proteins / Q000032:analysis* / Q000648:ultrastructure; D020543:Proteome / Q000032:analysis* / Q000648:ultrastructure; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity; D014978:Xenon
chemical_list: D029681:Arabidopsis Proteins; D004396:Coloring Agents; D005782:Gels; D010750:Phosphoproteins; D011506:Proteins; D020543:Proteome; D014978:Xenon