Acoustofluidic scanning fluorescence nanoscopy with a large field of view

Large-field nanoscale fluorescence imaging is invaluable for many applications,such as imaging subcellular structures,visualizing protein interactions,and high-resolution tissue imaging.Unfortunately,conventional fluorescence microscopy requires a trade-off between resolution and field of view due to the nature of the optics used to form the image.To overcome this barrier,we developed an acoustofluidic scanning fluorescence nanoscope that simultaneously achieves superior resolution,a large field of view,and strong fluorescent signals.The acoustofluidic scanning fluorescence nanoscope utilizes the superresolution capabilities of microspheres that are controlled by a programmable acoustofluidic device for rapid fluorescence enhancement and imaging.The acoustofluidic scanning fluorescence nanoscope resolves structures that cannot be resolved with conventional fluorescence microscopes with the same objective lens and enhances the fluorescent signal by a factor of~5 without altering the field of view of the image.The improved resolution realized with enhanced fluorescent signals and the large field of view achieved via acoustofluidic scanning fluorescence nanoscopy provides a powerful tool for versatile nanoscale fluorescence imaging for researchers in the fields of medicine,biology,biophysics,and biomedical engineering.

An acoustofluidic scanning nanoscope using enhanced image stacking and processing

Nanoscale optical resolution with a large field of view is a critical feature for many research and industry areas,such as semiconductor fabrication,biomedical imaging,and nanoscale material identification.Several scanning microscopes have been developed to resolve the inverse relationship between the resolution and field of view;however,those scanning microscopes still rely upon fluorescence labeling and complex optical systems.To overcome these limitations,we developed a dual-camera acoustofluidic nanoscope with a seamless image merging algorithm(alphablending process).This design allows us to precisely image both the sample and the microspheres simultaneously and accurately track the particle path and location.Therefore,the number of images required to capture the entire field of view(200×200μm)by using our acoustofluidic scanning nanoscope is reduced by 55-fold compared with previous designs.Moreover,the image quality is also greatly improved by applying an alpha-blending imaging technique,which is critical for accurately depicting and identifying nanoscale objects or processes.This dual-camera acoustofluidic nanoscope paves the way for enhanced nanoimaging with high resolution and a large field of view.