Spatially Resolved Genetic Analysis of Tissue Sections Enabled by Microscale Flow Confinement Retrieval and Isotachophoretic Purification. Issue 43 (16th September 2019)

• Record Type: Journal Article
• Authors: van Kooten, Xander F.; Petrini, Lorenzo F. T.; Kashyap, Aditya; Voith von Voithenberg, Lena; Bercovici, Moran; Kaigala, Govind V.
• Is Part Of: Angewandte Chemie international edition. Volume 58:Issue 43(2019)
• Page Start: 15259
• Page End: 15262
• Publication Date: 2019-09-16
• Subjects: cancer -- gene sequencing -- isotachophoresis -- tissue sections -- tumor heterogeneity; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201907150
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: 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. Abstract : Spatial sequencing : A method for spatially resolved genetic analysis of FFPE cell block and tissue sections is presented. This involves local sampling with hydrodynamic flow confinement of a lysis buffer, electrokinetic purification of nucleic acids from the lysate, and detection of single point mutations through amplification and sequencing.