Automated Dissection Protocol for Tumor Enrichment in Low Tumor Content Tissues.

Area of Science:

• Oncology
• Genomics
• Biotechnology

Background:

• Accurate tumor enrichment is crucial for reliable downstream genomic analysis, especially in low tumor content tissues.
• Traditional methods like macro-dissection and laser capture microdissection have limitations in precision, time, and cost.

Purpose of the Study:

• To introduce and evaluate an automated tissue dissection method for improved tumor enrichment in low tumor content tissues.
• To demonstrate the application of digital image annotation for precise targeting of tumor regions.

Main Methods:

• Utilizing digital hematoxylin and eosin (H&E) annotations to guide automated dissection of small tumor areas (250 µm²).
• Applying the method to unstained formalin-fixed paraffin-embedded (FFPE) or fresh frozen tissue sections (up to 20 µm thick).
• Automated harvesting of annotated regions for nucleic acid extraction and whole exome sequencing (WES).

Main Results:

• Automated dissection successfully enriches tumor content in low-content tissues, enabling reproducible data generation.
• The method improves accuracy and reproducibility while reducing the number of slides required.
• It allows for capture of pre- and post-harvest metrics, enhancing throughput and efficiency.

Conclusions:

• Automated tissue dissection with digital annotation provides an accurate, reproducible, and efficient solution for tumor enrichment.
• This methodology is applicable to both animal and human FFPE or fresh frozen tissues.
• The technique can be broadly used for region of interest enrichment to enhance sequencing applications in research and clinical settings.