Optimizing microlens arrays for incoherent HiLo microscopy

TL;DR

This paper introduces a cost-effective HiLo microscopy method using microlens arrays and incoherent LED light, demonstrating enhanced contrast through simulation and identifying optimal microlens parameters.

Contribution

It presents the first numerical study of MLA-based HiLo microscopy, showing how microlens parameters influence image contrast and quality.

Findings

MLA and incoherent light improve image contrast over traditional widefield microscopy.

MLA numerical aperture has minimal impact on HiLo image quality.

Optimal microlens pitch enhances contrast while avoiding artefacts.

Abstract

HiLo microscopy is a powerful, low-cost, and easily configurable technique for acquiring high-contrast optically-sectioned images. However, traditional HiLo microscopes are based on coherent light sources with diffusive glass plates or incoherent light sources with digital mirror devices (DMD) and spatial light modulators (SLM), which are more expensive. Here, we propose a new low-cost HiLo microscopy technique using MLAs and incoherent LED light sources. We simulated structured illumination (SI) patterns and HiLo image generation based on Fresnel diffraction and incoherent imaging. To observe how MLAs affect HiLo images, we used three common MLAs with specific microlens pitch and numerical aperture (NA) to generate periodic illumination patterns. According to our simulations, using MLAs and incoherent light sources can enhance the image contrast compared with a traditional widefield fluorescence microscope. We found that the MLA NA does not significantly affect HiLo images. A larger lens pitch can bring a higher image contrast. However, there is an optimized lens pitch. If the lens pitch is too high, artefacts are observed in HiLo images. To our knowledge, this is the first numerical study about MLA-based HiLo microscopy. This study can benefit researchers using MLAs and incoherent light sources to configure their low-cost HiLo microscopes.