Multiplexed and millimeter-scale superresolution imaging of cells and tissue sections via prism-illumination and microfluidics-enhanced DNA-PAINT

Abstract

AbstractSuperresolution microscopy (SRM) has been an enabling tool for biomedical research. A major limitation of SRM, however, is the small field-of-view (FOV), typically ~50μm x 50μm and up to ~200μm x 200μm in recent attempts, hampering its use in imaging large cell populations or clinical tissues. Here we report PRism-Illumination and Microfluidics-Enhanced DNA-PAINT (PRIME-PAINT) for efficient, multiplexed SRM across millimeter-scale FOVs. Unlike existing SRM, PRIME-PAINT uses prism-type illumination for robust DNA-PAINT with single FOVs up to a half millimeter. Through stitching, imaging over mm2 FOVs can be completed in under an hour per target. The on-stage microfluidics not only facilitates multiplexing but enhances image quality, particularly for tissue sections. We demonstrate the utility of PRIME-PAINT by analyzing ~106 caveolae structures in ~1,000 cells and imaging entire pancreatic cancer lesions from patient tissue biopsies. Thus, we expect PRIME-PAINT to be useful toward building multiscale, Google-Earth-like views of biological systems.