Lucid Workflow
Keeping the top-down workflow flowing.
Study Design
- Clearly define the scientific question
- Determine the optimal sample type(s) for study, considering potential sources of preanalytical bias
- Estimate the sample numbers necessary to achieve the required statistical power
System Qualification
- Optimize and monitor detector gain, laser stability, and laser energy
- Monitor mass accuracy and sensitivity
- Track instrument performance with Lucid™ proteomics software
Sample Preparation and ProteinChip® Array Binding
- Fractionate or enrich samples, as appropriate and feasible
- Apply crude samples or sample fractions to chromatographic arrays
- Capture and enrich proteins with specific biochemical properties on derivatized surfaces
Data Acquisition Set-Up
- Use virtual notebooks for sample tracking
- Create bioprocessor maps for sample layout
- Generate optimized AutoExecute methods for data collection
TOF Profiling Data Collection
- Collect data from each spot using automated Lucid rasterized firing pattern
- Detect peptides and proteins from 1.5 to 70 kD (linear mode)
- Import data into Lucid proteomics software
Analyze Data for Candidate Markers
- Use multiple modes of spectral visualization, as desired
- Detect peaks and generate peak clusters with automated detection tools
- Generate P value and receiver operator characteristic (ROC) statistics, trend plots, hierarchical clusters (heat maps), and/or principle component analysis (PCA) plots
Partial Purification and/or On-Chip Enrichment
- Screen chromatographic conditions with small volumes of sample on ProteinChip arrays
- Easily monitor target(s) purification on ProteinChip array of discovery
- Enrich target under 4 kD on chip or use SDS-PAGE as final purification step prior to passive elution and digestion
Collect TOF/TOF Fragmentation Data
- Collect TOF/TOF data from intact targets under 4 kD or larger MW target digests on chromatographic or normal phase ProteinChip arrays
Biomarker Identification (from Database Search)
- Export TOF/TOF data from Bruker’s flexAnalysis to BioTools and conduct MASCOT database search for target identification
Return to Top
