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See Also: | Flamingo Fluorescent Gel Stain | PDQuest 2-D Analysis Software | Criterion Precast Gel System |

The EXQuest spot cutter is a precision instrument that accurately locates and excises protein bands or spots from one- and two-dimensional gels or blots. It then loads them into microplates for downstream processing and analysis. It is designed to cut spots or bands from gels and blots with high efficiency, then deliver them to 96- and 384-well microplates or 96-tube racks. The EXQuest spot cutter is designed to be both robust in construction and simple to operate, with the capability to image gels and blots that are either visibly stained (for example, with silver or Coomassie Blue) or fluorescently stained (with SYPRO Ruby or Flamingo Stain).

The EXQuest spot cutter is a fully integrated system controlled through PDQuest 2-D analysis software or Quantity One 1-D analysis software. It consists of an enclosure, an imaging system, a fluidics system, robotics, sensors, a cutting head, a gel tray, a microplate rack, and a wash station.

EXQuest Gel Excision Versatility

The EXQuest spot cutter allows you to use any of the common proteome separation and staining methods:

• Freestanding 2-D and 1-D SDS-PAGE gels
• 2-D and 1-D gels cast on plastic or glass backing
• PVDF and nitrocellulose membrane blots
• Coomassie, silver, and other visible stains
• Flamingo fluorescent gel stain, SYPRO Ruby protein gel stain, and other fluorescent stains

The EXQuest Spot Cutter Provides Big Performance on the Bench

• Cuts 600 spots per hour
• System accuracy of ±0.1 mm ensures accurate excision of even small, closely grouped spots
• Reduces manual labor and improves throughput of protein spots for analysis
• Error-free data capture, from image analysis to mass spectrometry
• Accommodates any commercially available large or mini format gel or PVDF membrane blot in a small bench footprint
• Built-in wash wells provide customized tip washing between cuts for extra contamination control
• PDQuest software automatically selects spots in order from lowest to highest amount of protein, minimizing the chance of carryover contamination from spot to spot
• Quantity One software provides convenient tools for 1-D gel cutting
• Cutting tip produces precise cuts without damage or distortion, on gels or PVDF membranes

Fluorescence Detection Capability

The EXQuest spot cutter enables fluorescent dyes, such as Flamingo and SYPRO Ruby, to be used in the process of protein spot cutting. The enclosure acts as a darkroom with UV lights for the spot cutter, and is coordinated through the software during image acquisition.

• Highly accurate spot cutting from gels or membranes stained with visible or fluorescent protein stains, such as Flamingo and SYPRO Ruby
• Quick-change settings to switch between UV and visible light modes
• A small bench footprint that fits into any proteomics laboratory
• Compatibility with ProteomeWorks and ProteomeWorks Plus spot cutters already in use worldwide

For 2-D gel analysis, PDQuest software and the EXQuest spot cutter are integrated so that image analysis sets that reveal interesting proteins are used to direct automated spot cutting. Not only is the spot selection more directed, but the tracking of spot information is also controlled, from image analysis to protein annotation.

The EXQuest spot cutter and PDQuest software are fully integrated.

PDQuest image analysis software analyzes complex 2-D gel patterns and makes quantitative and qualitative comparisons over large sets of experimental data to discover protein changes. The analyses are compiled into matchsets (sets of gels that have been grouped together for the purpose of spot comparison), which are used to direct automated spot excision. This integration provides a valuable improvement in workflow and data tracking over other spot-cutting methods. PDQuest software tracks each protein spot, through spot cutting to protein identification.

The integration of PDQuest and Quantity One software with the EXQuest spot cutter provides:

• The ability to automatically cut bands or lanes from 1-D gels (example 1)
• The ability to resolve overlapped spots and cut them as 2 separate spots with the precision cutting mechanism, allowing excision and analysis of closely migrating spots (example 2)
• Improved workflow using spot-cut selections from analysis sets created in PDQuest
• Simple and accurate software-directed gel alignment on the spot cutter
• Spot tracking from gel to microplate location, simplifying data tracking and protein annotations
• Automatic calculation of the number of cuts that can be made from large individual protein spots; this ensures that as much protein from each spot as possible is automatically harvested into a single well in the microplate (example 3 and example 4)
• Spot review options for quick viewing of relative intensity levels (example 5)
• Export of spot-cut list to a spreadsheet to track data with any type of protein identification method (example 6)
• Confirmation of cuts post-excision (example 7)

For more information, visit the Expression Proteomics web site.