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Spatially Resolved Genetic Analysis of Tissue Sections Enabled by Microscale Flow Confinement Retrieval and Isotachophoretic Purification
Author(s) -
Kooten Xander F.,
Petrini Lorenzo F. T.,
Kashyap Aditya,
Voith von Voithenberg Lena,
Bercovici Moran,
Kaigala Govind V.
Publication year - 2019
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201907150
Subject(s) - microscale chemistry , lysis , genotyping , nucleic acid , kras , chemistry , microbiology and biotechnology , biology , mutation , genotype , biochemistry , mathematics education , mathematics , gene
We have developed a method for spatially resolved genetic analysis of formalin‐fixed paraffin‐embedded (FFPE) cell block and tissue sections. This method involves local sampling using hydrodynamic flow confinement of a lysis buffer, followed by electrokinetic purification of nucleic acids from the sampled lysate. We characterized the method by locally sampling an array of points with a circa 200 μm diameter footprint, enabling the detection of single KRAS and BRAF point mutations in small populations of RKO and MCF‐7 FFPE cell blocks. To illustrate the utility of this approach for genetic analysis, we demonstrate spatially resolved genotyping of FFPE sections of human breast invasive ductal carcinoma.