A 'waterfall' transfer-based workflow for improved quality of tissue microarray construction and processing in breast...

Area of Science:

• Oncology
• Biomarker Discovery
• Pathology

Background:

• Identifying cancer biomarkers is crucial for early diagnosis, treatment selection, and prognosis.
• Tissue microarrays (TMAs) are vital for high-throughput analysis of clinical samples.
• Challenges exist in constructing and processing TMAs for reliable research.

Purpose of the Study:

• To present a robust workflow for constructing high-density breast cancer TMAs.
• To overcome obstacles in TMA construction and processing for high-quality research sections.
• To ensure sample integrity for subsequent immunohistochemistry (IHC) and immunofluorescence (IF) staining.

Main Methods:

• Constructed high-density breast cancer TMAs using 406 tissue cores from 245 patients.
• Utilized a rotary microtome with an automated transfer system for sectioning.
• Performed IHC and IF staining for nine proteins, manually assessing core quality and quantity.

Main Results:

• Achieved 97.1% core recovery after sectioning.
• Maintained 96% evaluability of remaining cores after staining.
• Observed higher loss rates in normal tissue cores compared to tumor cores.

Conclusions:

• The developed workflow enables the robust manufacturing of high-density breast cancer TMAs.
• This method minimizes sample loss, ensuring high-quality sections for IHC and IF.
• The workflow supports reliable biomarker validation in cancer research.