Abstract
Integrated multimodal cross-sectional or volumetric imaging techniques give us fruitful information to understand the behavior or status of target objects such as biological samples. Most of the reported systems for this purpose are either time consuming due to scanning or use additional reference beams such as in interferometry. Therefore, fast, simple, highly efficient, and powerful multimodal imaging systems that can perform cross-sectional imaging with simple algorithms are worth to be investigated. In this paper, a multimodal technique for cross-sectional quantitative phase and fluorescence imaging with computational microscopy is presented. We combine cross-sectional fluorescence and quantitative phase imaging by using the transport of intensity equation (TIE) and numerical wave propagation. The amplitude and phase of the fluorescence light wave with partially spatial coherence are obtained from three defocused intensity patterns. The proposed hybrid imaging system is simple, compact, and non-iterative. We present experimental results of microbeads and fluorescent protein-labeled living cells of the moss Physcomitrella patens to demonstrate the performance of the proposed imaging system.
Highlights
INTRODUCTIONF LUORESCENCE imaging enables the visualization of the functional details of samples by labeling certain molecules
F LUORESCENCE imaging enables the visualization of the functional details of samples by labeling certain moleculesManuscript received October 8, 2020; revised February 5, 2021; accepted March 1, 2021
To perform the experiment for the demonstration of simultaneous cross-sectional fluorescence and quantitative phase imaging, the optical setup shown in Fig. 1 is used
Summary
F LUORESCENCE imaging enables the visualization of the functional details of samples by labeling certain molecules. Several techniques such as optical interferometry [18], single-pixel imaging and wavefront sensing [19], [20], ptychography [21] and transport of intensity equation imaging (TIE) [22]–[28], have been utilized for the visualization of phase information Most of these techniques have been combined with fluorescence imaging to get additional functional information of the biological sample to be imaged [29]–[37]. LED2 is used to illuminate the sample In this case, the light passes through the same components as in the case of fluorescence up to DM2, minimizing the requirement of components unlike other multimodal systems performing interferometric or holography based multimodal imaging.
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