Fast Inertia-Free Volumetric Light-Sheet Microscope
Fast noninvasive three-dimensional (3D) imag-ing is crucial for quantitatively studying highly dynamic events ranging from flow cytometry to developmental biology. Light-sheet microscopy has emerged as the tool-of-choice for 3D characterization of rapidly evolving systems. However, to obtain a 3D image, either the sample or parts of the microscope are moved, limiting the acquisition speed. Here, we propose a novel inertia-free light-sheet-based scheme for volumetric imaging at high temporal resolution. Our approach comprises a novel combination of an acousto-optic scanner to produce tailored illumination and an acoustic-optofluidic lens, placed in the detection path to provide extended dept…
Three-dimensional multiple-particle tracking with nanometric precision over tunable axial ranges
The precise localization of nanometric objects in three dimensions is essential to identify functional diffusion mechanisms in complex systems at the cellular or molecular level. However, most optical methods can achieve high temporal resolution and high localization precision only in two dimensions or over a limited axial (z) range. Here we develop a novel wide-field detection system based on an electrically tunable lens that can track multiple individual nanoscale emitters in three dimensions over a tunable axial range with nanometric localization precision. The optical principle of the technique is based on the simultaneous acquisition of two images with an extended depth of field while …
A Novel Fast Volumetric Light Sheet Microscopy
Fast noninvasive three-dimensional (3D) imaging is crucial for the quantitative understanding of highly dynamic biological processes. Over the last decades, several fluorescence microscopy techniques have been developed in order to provide a faster and deeper imaging of thick biological samples [1]. Within this framework, Light Sheet Fluorescence Microscopy (LSFM) has emerged as a powerful imaging tool for 3D imaging of thick samples ranging from single cells to entire animals [2,3].However, to obtain a 3D reconstruction either sample or microscope parts usually need to be moved limiting the acquisition speed and inducing possible interferences in volume recording. To solve this problem, he…