Optical Sectioning Microscopy Through Single-Shot Lightfield Protocol

E Sánchez-Ortiga,G Saavedra,G Scrofani,M Martinez-Corral

doi:10.1109/access.2020.2966323

E Sánchez-Ortiga, G Saavedra + Show 2 more

Open Access

https://doi.org/10.1109/access.2020.2966323

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Abstract

Optical sectioning microscopy is usually performed by means of a scanning, multi-shot procedure in combination with non-uniform illumination. In this paper, we change the paradigm and report a method that is based in the lightfield concept, and that provides optical sectioning for 3D microscopy images after a single-shot capture. To do this we first capture multiple orthographic perspectives of the sample by means of Fourier-domain integral microscopy (FiMic). The second stage of our protocol is the application of a novel refocusing algorithm that is able to produce optical sectioning in real time, and with no resolution worsening, in the case of sparse fluorescent samples. We provide the theoretical derivation of the algorithm, and demonstrate its utility by applying it to simulations and to experimental data.

Highlights

In the past few decades integral imaging has been proven to be a successful alternative to conventional photography [1]–[7]
We report the results of lightfield experiments, using different 3D fluorescent samples, which demonstrate the utility of our method
SIMULATION We performed a set of simulations in which, by means of Eq(1), we calculated the field intensity captured by a Fourier-domain integral microscopy (FiMic)

Summary

INTRODUCTION

In the past few decades integral (or lightfield) imaging has been proven to be a successful alternative to conventional photography [1]–[7]. The axial position of a given emitter can be measured by means of the relative angle of the point spread function generated in the image plane. The position of different individual emitters is localized at different times, even though 3D information can be obtained by means of a single-shot, these technique require thousands of realizations in order to provide a single 3D image of the sample, which make them unfeasible for real-time acquisition Note that, all these techniques require the measurement of external parameters: relative intensity of two detectors and the axial response, phase-shift of the self-interference pattern and angle of the aberrated point spread function. Mz m where Im(x) is the irradiance distribution on the m-th EI

THE STANDARD RECONSTRUCTION ALGORITHM

RESULTS

GPU ACCELERATED ALGORITHM

CONCLUSION