DeCyder differential analysis with statistical confidence

DeCyder™ (Amersham Biosciences/GE) has been specifically developed as a key element of the Ettan™ DIGE system. This proprietary software significantly increases throughput by accurately addressing measurement of protein differences with statistical confidence and reducing hands-on time from days to minutes with minimal user-to-user variation. DeCyder automatically detects, matches and analyzes protein spots in multiplexed fluorescent images, and is able to give routine detection of < 10% differences with > 95 % confidence. Statistical analysis is carried out on each and every difference.
Pictures courtesy of GE
DeCyder consists of two analysis modules: 1. Differential In-gel Analysis (DIA) The DIA software module provides consistent and accurate co-detection of image pairs from the same gel using novel algorithms. Background subtraction, quantitation, normalization and first level matching within gel are automated for high-throughput analysis with low experimental variation.

DIA output:

  • Co-detection of image pairs
  • Automated detection, background subtraction
  • Quantitation, normalization and first level (In-gel) matching
  • Low user interaction, high throughput
  • Very low experimental variation
  • Optimized workflow for 2-D DIGE analysis
2. Biological Variation Analysis (BVA) Using batch-processing, up to several hundred gel pair-images can be processed unattended for significant time savings. Images are then matched between gels using the BVA software module that looks for consistency of differences between samples across all the gels and applies statistics to associate a level of confidence for each of those differences. Finally, the XML Toolbox can be used to export spot data from DIA or BVA for downstream analysis. BVA output:
  • Automated gel-to-gel matching
  • Statistical / Trend analysis
Internal standardization Appropriate Experimental Design and Statistical Analysis (EDSA) is central to 2-D DIGE technology and ensures the best possible results. The recommended EDSA utilizes the internal standard already described in the CyDye DIGE Fluors section, to eliminate gel-to-gel variation. Automatic statistical analysis 2-D DIGE is the only electrophoresis technology that enables internal standardization of differences in protein abundance levels. DeCyder Differential Analysis Software has an EDSA setting feature for fully automated statistical analysis. The DeCyder Differential Analysis Software automatically generates a list of statistically significant differences, based on the threshold parameters determined by the user.
  • DeCyder Differential Analysis Software: the facts
  • Automatic analysis: significantly reduces hands-on time
  • No image editing: results are objective
  • No spot matching within gel: eliminates errors
  • Spot statistics are automatically presented
  • Designed to use an internal standard: increases accuracy and simplifies gel-to-gel matching