Enhanced quantitative phase imaging in self-interference digital holographic microscopy using an electrically focus tunable lens

Schubert, Robin; Vollmer, Angelika; Ketelhut, Steffi; Kemper, Björn

doi:10.1364/boe.5.004213

Cited by 31 publications

(11 citation statements)

References 38 publications

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“…Neurons exhibit growth rates an order of magnitude smaller than adherent cells, which, due to mitosis, double in mass, roughly, every 10-40 hours (see for points of comparison [55, 56]). In practice, this introduces an instrument sensitivity requirement that is difficult to meet with many methods as imaging artifacts often exceed the 0.15 Rad phase shift of a typical neurite[12, 57]. Comparing to limited previous efforts, the values reported in this work are comparable with those in[58], within variability due to cell type and confluence.…”

Section: Resultsmentioning

confidence: 71%

Real-time halo correction in phase contrast imaging

Kandel

Fanous

Best-Popescu

et al. 2017

Preprint

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As a label-free, nondestructive method, phase contrast is by far the most popular microscopy technique for routine inspection of cell cultures. Yet, features of interest such as extensions near cell bodies are often obscured by a glow, which came to be known as the halo. Advances in modeling image formation have shown that this artifact is due to the limited spatial coherence of the illumination. Yet, the same incoherent illumination is responsible for superior sensitivity to fine details in the phase contrast geometry. Thus, there exists a trade-off between high-detail (incoherent) and low-detail (coherent) imaging systems. In this work, we propose a method to break this dichotomy, by carefully mixing corrected low-frequency and high-frequency data in a way that eliminates the edge effect. Specifically, our technique is able to remove halo artifacts at video rates, requiring no manual interaction or a priori point spread function measurements. To validate our approach, we imaged standard spherical beads, sperm cells, tissue slices, and red blood cells. We demonstrate the real-time operation with a time evolution study of adherent neuron cultures whose neurites are revealed by our halo correction. We show that with our novel technique, we can quantify cell growth in large populations, without the need for thresholds and calibration.

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Section: Resultsmentioning

confidence: 71%

Real-time halo correction in phase contrast imaging

Kandel

Fanous

Best-Popescu

et al. 2017

Preprint

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“…Neurons exhibit growth rates an order of magnitude smaller than adherent cells, which, due to mitosis, double in mass, roughly, every 10-40 hours (see for points of comparison [24,64]). In practice, this introduces an instrument sensitivity requirement that is difficult to meet with many methods as imaging artifacts often exceed the 0.15 Rad phase shift of a typical neurite [12,65]. Comparing to limited previous efforts, the values reported in this work are comparable with those in reference [66], within variability due to cell type and confluence.…”

Section: Resultsmentioning

confidence: 71%

Real-time halo correction in phase contrast imaging

Kandel¹,

Fanous²,

Best-Popescu³

et al. 2018

Biomed. Opt. Express

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As a label-free, nondestructive method, phase contrast is by far the most popular microscopy technique for routine inspection of cell cultures. However, features of interest such as extensions near cell bodies are often obscured by a glow, which came to be known as the . Advances in modeling image formation have shown that this artifact is due to the limited spatial coherence of the illumination. Nevertheless, the same incoherent illumination is responsible for superior sensitivity to fine details in the phase contrast geometry. Thus, there exists a trade-off between high-detail (incoherent) and low-detail (coherent) imaging systems. In this work, we propose a method to break this dichotomy, by carefully mixing corrected low-frequency and high-frequency data in a way that eliminates the edge effect. Specifically, our technique is able to remove halo artifacts at video rates, requiring no manual interaction or point spread function measurements. To validate our approach, we imaged standard spherical beads, sperm cells, tissue slices, and red blood cells. We demonstrate real-time operation with a time evolution study of adherent neuron cultures whose neurites are revealed by our halo correction. We show that with our novel technique, we can quantify cell growth in large populations, without the need for thresholds and system variant calibration.

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“…The second strategy is based on optical instrumentation development. 18 , 19 As far as QP images in a microscopy setting are concerned, efficient coherent noise reduction approaches based on either object or camera motion, or on the use of different laser modes have been developed. 20 – 22 Specifically, in a transmission configuration aimed at exploring biological cells, a lateral shift of the camera 21 significantly reduced the coherent noise, while a noise reduction approaching the ideal curve of

was obtained by slightly moving the object.…”

Section: Introductionmentioning

confidence: 99%

Polychromatic digital holographic microscopy: a quasicoherent-noise-free imaging technique to explore the connectivity of living neuronal networks

Larivière-Loiselle

Bélanger²,

Marquet³

2020

Neurophoton.

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Significance: Over the past decade, laser-based digital holographic microscopy (DHM), an important approach in the field of quantitative-phase imaging techniques, has become a significant label-free modality for live-cell imaging and used particularly in cellular neuroscience. However, coherent noise remains a major drawback for DHM, significantly limiting the possibility to visualize neuronal processes and precluding important studies on neuronal connectivity. Aim: The goal is to develop a DHM technique able to sharply visualize thin neuronal processes. Approach: By combining a wavelength-tunable light source with the advantages of hologram numerical reconstruction of DHM, an approach called polychromatic DHM (P-DHM), providing OPD images with drastically decreased coherent noise, was developed. Results: When applied to cultured neuronal networks with an air microscope objective (20×, 0.8 NA), P-DHM shows a coherent noise level typically corresponding to 1 nm at the single-pixel scale, in agreement with the 1∕ ffiffiffiffi N p-law, allowing to readily visualize the 1-μm-wide thin neuronal processes with a signal-to-noise ratio of ∼5. Conclusions: Therefore, P-DHM represents a very promising label-free technique to study neuronal connectivity and its development, including neurite outgrowth, elongation, and branching.

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Enhanced quantitative phase imaging in self-interference digital holographic microscopy using an electrically focus tunable lens

Cited by 31 publications

References 38 publications

Real-time halo correction in phase contrast imaging

Real-time halo correction in phase contrast imaging

Real-time halo correction in phase contrast imaging

Polychromatic digital holographic microscopy: a quasicoherent-noise-free imaging technique to explore the connectivity of living neuronal networks

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