Microdissection Methods Utilizing Single-Cell Subtype Analysis and the Impact on Precision Medicine

Methods Mol Biol. 2022:2394:93-107. doi: 10.1007/978-1-0716-1811-0_7.

Abstract

Improving the utilization of tumor tissue from diagnostic biopsies is an unmet medical need. This is especially relevant today in the rapidly evolving precision oncology field where tumor genotyping is often essential for the indication of many advanced and targeted therapies. National Comprehensive Cancer Network (NCCN) guidelines now mandate molecular testing for clinically actionable targets in certain malignancies. Utilizing advanced stage lung cancer as an example, an improved genotyping approach for solid tumors is possible. The strategy involves optimization of the microdissection process and analysis of a large number of identical target cells from formalin-fixed paraffin-embedded (FFPE) specimens sharing similar characteristics, in other words, single-cell subtype analysis. The shared characteristics can include immunostaining status, cell phenotype, and/or spatial location within a histological section. Synergy between microdissection and droplet digital PCR (ddPCR) enhances the molecular analysis. We demonstrate here a methodology that illustrates genotyping of a solid tumor from a small tissue biopsy sample in a time- and cost-efficient manner, using immunostain targeting as an example.

Keywords: Biomarker; Cancer; Droplet digital PCR (ddPCR); Laser capture microdissection (LCM); Lung cancer; Precision medicine; Precision oncology; Single-cell analysis; Tumor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Formaldehyde
  • Humans
  • Microdissection* / methods
  • Neoplasms*
  • Paraffin Embedding / methods
  • Polymerase Chain Reaction / methods
  • Precision Medicine
  • Tissue Fixation / methods

Substances

  • Formaldehyde