Fluholoscopy. Compact and Simple Platform Combining Fluorescence and Holographic Microscopy

TL;DR

This paper introduces a simple, cost-effective microscope platform capable of simultaneous fluorescence and phase imaging in a single snapshot, enhancing biomedical analysis with minimal complexity.

Contribution

It presents a novel, compact design that combines fluorescence and holographic microscopy using a single wavelength and dual cameras, enabling real-time dual-mode imaging.

Findings

Successfully performed calibration and independent analysis of both imaging modes.

Validated the platform with static and dynamic biological samples.

Demonstrated real-time imaging of flowing and live specimens.

Abstract

The combination of different imaging modalities into single imaging platforms has a strong potential in biomedical sciences since it permits the analysis of complementary properties of the target sample. Here, we report on an extremely simple, cost-effective, and compact microscope platform for achieving simultaneous fluorescence and quantitative phase imaging modes with the capability of working in a single snapshot. It is based on the use of a single illumination wavelength to both excite the sample fluorescence and provide coherent illumination for phase imaging. After passing the microscope layout, the two imaging paths are separated by using a bandpass filter and the two imaging modes are simultaneously obtained by using two digital cameras. We first present calibration and analysis of both fluorescence and phase imaging modalities working independently and, later on, experimental validation for the proposed common-path dual-mode imaging platform considering static (resolution test targets, fluorescent micro-beads and water-suspended lab-made cultures) as well as dynamic (flowing fluorescent beads, human sperm cells and live specimens from lab-made cultures) samples.