High-Definition Imaging Software (HDI) 1.7 — New Features And Benefits

Significance of the Topic

Mass spectrometry imaging (MSI) has become a critical technique in analytical chemistry, enabling spatial mapping of molecular species directly from sample surfaces. HDI software facilitates streamlined workflows for both MALDI and DESI imaging modalities, providing researchers and quality control analysts with tools to visualize, quantify, and interpret complex datasets. Enhancements in HDI 1.7 improve sensitivity, specificity, and throughput, addressing key challenges in targeted and discovery-based imaging studies.

Study Objectives and Overview

This release aims to introduce new features in HDI 1.7 that expand capabilities for targeted workflows, quantitative analysis, ROI management, and mass accuracy correction. The software integration across MassLynx, EZ Info, and MicroApps supports a comprehensive end-to-end MSI pipeline from data acquisition to advanced statistical analysis.

Methodology and Workflow

HDI 1.7 supports multiple acquisition modes such as SONAR, HDMS, MRM, SIR, and MS/MS. The targeted workflow leverages the DESI XS TQ Method Editor on the Xevo TQ Absolute platform to monitor up to 32 ion transitions simultaneously, automatically extracting and visualizing ion images for scheduled functions. The MSI Quantify MicroApp enables ROI selection, calibration curve generation, and concentration calculations for both known standards and unknown analytes.

Used Instrumentation

• Waters HDI 1.7 software suite
• MassLynx data acquisition and processing environment
• EZ Info for statistical analyses
• MS Imaging MicroApps (quantification and segmentation)
• Xevo TQ Absolute triple quadrupole mass spectrometer
• DESI XS TQ Method Editor for targeted MRM/SIR workflows

Main Results and Discussion

HDI 1.7 delivers enhanced targeted imaging, demonstrating high specificity and sensitivity in MRM and SIR experiments. The MSI Quantify MicroApp streamlines quantitative assessments, producing reliable calibration curves and concentration maps. ROI save and reload features improve workflow reproducibility; exports now preserve individual pixel spectra and dimensions for detailed downstream processing. The new Continuous Lock Mass Correction routine maintains mass accuracy throughout long acquisitions, resulting in smoother peak alignment and improved spectral fidelity.

Benefits and Practical Applications

• Accelerated targeted MSI workflows with up to 32 simultaneous transitions
• End-to-end quantitation capabilities directly within imaging datasets
• Improved ROI management for reproducible mask-based processing
• Enhanced mass accuracy through continuous lock mass correction
• Flexible spectral viewing options, including average, summed, and maximum intensity displays
• Support for high-precision mass measurements to five decimal places

Future Trends and Potential Applications

Advancements in MSI software will likely focus on machine learning–driven image segmentation, real-time quantitative feedback, and integration with multi-omics data. Enhanced cloud-based processing and collaborative platforms may further democratize access to high-definition imaging analysis. Continued development of targeted and untargeted imaging will expand applications in pharmaceutical research, clinical diagnostics, and spatial omics.

Conclusion

HDI 1.7 enhances Waters’ MS imaging platform by delivering powerful targeted workflows, robust quantitation tools, and improved data management features. These developments enable researchers to achieve higher confidence in spatially resolved molecular analyses, paving the way for broader adoption of MSI in both research and applied settings.