Optimizing Tissue Preservation for High-Resolution Confocal Imaging of Single-Molecule RNA-FISH.

ABSTRACT: Over the past century, formalin-fixed, paraffin-embedded (FFPE) tissue samples have represented the standard for basic histology and immunostaining. However, FFPE has several limitations and less stringent tissue preservation methods are required for the visualization of nucleic acids at high resolution, particularly those that are expressed at low levels. Here, we describe the FFPE properties that negatively impact RNA integrity, an alternative tissue preservation technique that prevents RNA loss, and the steps necessary to optimize slide preparation for single-molecule RNA fluorescent in situ hybridization (smRNA-FISH) and imaging by confocal microscopy. This strategy retains RNA quality and eliminates formalin-induced artifacts, thereby producing high-resolution, diffraction-limited con

SUBMITTER: Redmayne N 

PROVIDER: S-EPMC7202135 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature