A plane wave analysis of coherent holographic image reconstruction by phase transfer

TL;DR

This paper introduces a mathematical model and experimental validation for CHIRPT, a novel fluorescent imaging technique combining light-sheet illumination with non-imaging detection to improve speed and scattering robustness in biological imaging.

Contribution

The paper develops a formal mathematical framework for CHIRPT imaging under coherent illumination and provides experimental verification of the model.

Findings

Mathematical formalism for CHIRPT presented

Experimental data confirms theoretical predictions

CHIRPT enhances imaging speed and scattering robustness

Abstract

Fluorescent imaging plays a critical role in a myriad of scientific endeavors, particularly in the biological sciences. Three-dimensional imaging of fluorescent intensity often requires serial data acquisition, that is voxel-by-voxel collection of fluorescent light emitted throughout the specimen with a non-imaging single-element detector. While non-imaging fluorescence detection offers some measure of scattering robustness, the rate at which dynamic specimens can be imaged is severely limited. Other fluorescent imaging techniques utilize imaging detection to enhance collection rates. A notable example is light-sheet fluorescence microscopy, also known as selective-plane illumination microscopy (SPIM), which illuminates a large region within the specimen and collects emitted fluorescent light at an angle either perpendicular or oblique to the illumination light sheet. Unfortunately, scattering of the emitted fluorescent light can cause blurring of the collected images in highly turbid biological media. We recently introduced an imaging technique called coherent holographic image reconstruction by phase transfer (CHIRPT) that combines light-sheet-like illumination with non-imaging fluorescent light detection. By combining the speed of light-sheet illumination with the scattering robustness of non-imaging detection, CHIRPT is poised to have a dramatic impact on biological imaging, particularly for $\textit{in vivo}$ preparations. Here we present the mathematical formalism for CHIRPT imaging under spatially coherent illumination and present experimental data that verifies the theoretical model.