Quantitative phase imaging of biological cells using spatially low and temporally high coherent light source

Ahmad, Azeem; Dubey, Vishesh; Singh, Gyanendra; Singh, Veena; Mehta, Dalip Singh

doi:10.1364/ol.41.001554

Cited by 50 publications

(45 citation statements)

References 17 publications

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“…Unlike to SIM microscopy, this technique does not require any labelling agent to quantify the morphological features [33]. The details of the set up can be found elsewhere [24,26,27]. A 60 × microscope objective (NA = 1.2) is used for the imaging of the control and inflamed live cells.…”

Section: Quantitative Phase Imaging Of Macrophages and Trophoblastsmentioning

confidence: 99%

“…In label-free optical imaging, quantitative phase microscopy (QPM) is a popular technique for the quantitative analysis of live biological specimens [24]. QPM techniques are capable to quantify various parameters associated with biological specimen, such as cell dynamics (fluctuations in cell thickness and/or RI), cell morphology and cell dry mass (non-aqueous content) [24][25][26][27]. Digital holographic microscopy (DHM) is one of the interferometric QPM techniques to extract the quantitative information of the live cell [26].…”

Section: Introductionmentioning

confidence: 99%

“…Digital holographic microscopy (DHM) is one of the interferometric QPM techniques to extract the quantitative information of the live cell [26]. It is very useful technique for the single cell analysis with ease of operation and compatibility [27,28]. In connection with vigorous algorithms of numerical reconstruction of interferograms for data measurement, DHM provides a fascinating window in modern microscopy for quantitative analysis of the specimen.…”

Section: Introductionmentioning

confidence: 99%

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Quantitative assessment of morphology and sub-cellular changes in macrophages and trophoblasts during inflammation

Singh

Dubey

Wolfson

et al. 2020

Preprint

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During inflammatory condition in pregnancy, the macrophages present at the feto-maternal junction release an increased amount of NO and pro-inflammatory cytokines such as TNF-α and INF-γ, which can disturb the trophoblast functions and thereby the pregnancy outcome. Measurement of the cellular and subcellular morphological modifications associated with inflammatory responses are important in order to quantify the extent of trophoblast dysfunction for clinical implication. With this motivation, we investigated morphological, cellular and sub-cellular changes in externally inflamed RAW264.7 (macrophage) and HTR-8/SVneo (trophoblast) using structured illumination microscopy (SIM) and quantitative phase microscopy (QPM). We monitored the production of nitric oxide (NO), changes in cell membrane and mitochondrial structure of macrophages and trophoblasts when exposed to different concentration of pro-inflammatory agents (LPS and TNF-α). In vitro NO production by LPS-induced macrophages increased 22-folds as compared to controls, whereas no significant NO production was seen after TNF-α challenge. Under similar conditions as with macrophages, trophoblasts did not produce NO following either LPS or TNF-α challenge. Super-resolution SIM imaging showed changes in the morphology of mitochondria and plasma membrane in macrophages following LPS challenge and in trophoblasts following TNF-α challenge. Label-free QPM showed a decrease in the optical thickness of the LPS-challenged macrophages while TNF-α having no effect. The vice-versa is observed for the trophoblasts. We further exploited machine learning approaches on QPM dataset to detect and to classify the inflammation with an accuracy of 99.9% for LPS-challenged macrophages and 98.3% for TNF-αchallenged trophoblasts. We believe that the multi-modal advanced microscopy methodologies coupled with machine learning approach could be an alternative way for early detection of pregnancy related inflammation after clinical studies.

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Section: Quantitative Phase Imaging Of Macrophages and Trophoblastsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Quantitative assessment of morphology and sub-cellular changes in macrophages and trophoblasts during inflammation

Singh

Dubey

Wolfson

et al. 2020

Preprint

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“…aforementioned issues with the conventional light sources 3,7,9,10 . PTS can be generated by employing different approaches such as rotating diffuser, vibrating multiple multi-mode fiber bundle (MMFB) and electro-active-polymer rotational micro-optic diffuser 7,[11][12][13] .…”

Section: Introductionmentioning

confidence: 99%

“…PTS have been successfully implemented in optical profilometry, quantitative phase microscopy and OCM/T techniques to improve their performance 3,7,9 . In the past, a lot of theoretical and experimental work has also been done to understand and study the coherence properties of PTS. The coherence properties, especially the longitudinal spatial coherence (LSC) properties, of PTS as a function of source size, numerical aperture (NA) of microscope objective and wavelength have been studied previously 9,11,12,14,15 .…”

Section: Introductionmentioning

confidence: 99%

Relationship between the source size at the diffuser plane and the longitudinal spatial coherence function of the optical coherence microscopy system

Usmani¹,

Ahmad²,

Joshi³

et al. 2019

J. Opt. Soc. Am. A

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Coherence properties and wavelength of light sources are indispensable for optical coherence microscopy/tomography (OCM/T) as they greatly influence the signal to noise ratio, axial resolution, and penetration depth of the system. In the present letter, we investigated the longitudinal spatial coherence properties of pseudo/thermal light source (PTS) as a function of spot size at the diffuser plane, which is controlled by translating microscope objective lens towards or away from the diffuser plane. The axial resolution of PTS is found to be maximum ~ 13 µm for the beam spot size of 3.5 mm at the diffuser plane. The change in the axial resolution of the system as the spot size is increased at the diffuser plane is further confirmed by performing experiments on standard gauge blocks of height difference of 15 µm. Thus, by appropriately choosing the beam spot size at the diffuser plane, any monochromatic laser light source depending on the biological window can be utilized to obtain high axial-resolution with large penetration depth and speckle free tomographic images of multilayered biological specimens irrespective of the source's temporal coherence length. In addition, PTS could be an attractive alternative light source for achieving high axial-resolution without needing chromatic aberration corrected optics and dispersion-compensation mechanism unlike conventional setups.Recently, a number of studies have been put forward to synthesize a spatially extended and temporally coherent light source named pseudo-thermal light source (PTS) to overcome

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Coherence‐Tailored Multiwavelength High‐Speed Quantitative Phase Imaging with a High Phase Stability via a Frequency Comb

Boonruangkan

Farrokhi

Rohith

et al. 2020

Advanced Photonics Research

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Coherent imaging enables noninvasive, label‐free, and quantitative monitoring of the dynamic motions of transparent microobjects requested in life sciences, biochemistry, material sciences, and fluid mechanics. Quantitative phase imaging (QPI), a coherent imaging technique, provides full‐field optical phase information through light interference. The use of coherence, however, inevitably accompanies phase ambiguity and coherent artifacts, such as speckle, diffraction, and parasitic interference, which severely deteriorate the interferograms to hinder successful phase reconstruction. Herein, it is demonstrated that a frequency comb can newly provide a wide coherence tunability for higher visibility interferograms, phase‐coherent multiple wavelengths for extracting physical height information from refractive index, and higher phase stability (2.39 × 10−3 at 10 s averaging time) at a higher speed up to 16.9 kHz. These superior characteristics of frequency‐comb‐referenced QPI will enable in‐depth understanding of dynamic motions in cellular, biomolecular, and microphysical samples.

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Quantitative phase imaging of biological cells using spatially low and temporally high coherent light source

Cited by 50 publications

References 17 publications

Quantitative assessment of morphology and sub-cellular changes in macrophages and trophoblasts during inflammation

Quantitative assessment of morphology and sub-cellular changes in macrophages and trophoblasts during inflammation

Relationship between the source size at the diffuser plane and the longitudinal spatial coherence function of the optical coherence microscopy system

Coherence‐Tailored Multiwavelength High‐Speed Quantitative Phase Imaging with a High Phase Stability via a Frequency Comb

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