Chip Based Nanoscopy: Towards Integration and High-throughput Imaging

Abstract

Super-resolution optical microscopy, commonly referred to as optical nanoscopy, has enabled imaging of biological samples with a resolution that was only achievable previously using electron microscopy. Optical nanoscopy is a rapidly growing field, with several different techniques and implementations that overcome the diffraction limit of light. However, the common nanoscope continues to be a rather complex, expensive and bulky instrument. Direct stochastic optical reconstruction microscopy (dSTORM) imaging was recently demonstrated using a waveguide platform for excitation in combination with a simple microscope for imaging. High refractive index waveguide materials have a high intensity evanescent field stretching around 100-200 nm outside the guiding material, which is ideally suited for total internal reflection fluorescence (TIRF) excitation over large areas. We demonstrate dSTORM imaging of the plasma membrane of liver sinusoidal endothelial cells (LSECs) and trophoblasts (HTR-8 cells) using waveguide excitation, with resolution down to around 70 nm. Additionally, we present TIRF imaging of LSEC micro-tubules over a 500 μm x 500 μm area, laying the foundation for large field of view (f-o-v) nanoscopy.