Quantitative Stain-Free and Continuous Multimodal Monitoring of Wound Healing In Vitro with Digital Holographic Microscopy

Bettenworth, Dominik; Lenz, Philipp; Krausewitz, Philipp; Brückner, Markus; Ketelhut, Steffi; Domagk, Dirk; Kemper, Björn

doi:10.1371/journal.pone.0107317

Cited by 81 publications

(52 citation statements)

References 61 publications

(80 reference statements)

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“…33 It has been shown that the measured phase in the image corresponds to the dry mass density distribution within the cell. 26,27 In contrast to existing DHM techniques, 4,7,18,19 the use of incoherent illumination enables high-quality quantitative phase imaging with strong suppression of coherent noise and parasitic interferences while providing high temporal stability and spatial uniformity of the phase measurement. 25 Using the approach described in Ref.…”

Section: Image Acquisitionmentioning

confidence: 99%

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Automated classification of cell morphology by coherence-controlled holographic microscopy

et al. 2017

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Abstract. In the last few years, classification of cells by machine learning has become frequently used in biology. However, most of the approaches are based on morphometric (MO) features, which are not quantitative in terms of cell mass. This may result in poor classification accuracy. Here, we study the potential contribution of coherence-controlled holographic microscopy enabling quantitative phase imaging for the classification of cell morphologies. We compare our approach with the commonly used method based on MO features. We tested both classification approaches in an experiment with nutritionally deprived cancer tissue cells, while employing several supervised machine learning algorithms. Most of the classifiers provided higher performance when quantitative phase features were employed. Based on the results, it can be concluded that the quantitative phase features played an important role in improving the performance of the classification. The methodology could be valuable help in refining the monitoring of live cells in an automated fashion. We believe that coherencecontrolled holographic microscopy, as a tool for quantitative phase imaging, offers all preconditions for the accurate automated analysis of live cell behavior while enabling noninvasive label-free imaging with sufficient contrast and high-spatiotemporal phase sensitivity.

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Section: Image Acquisitionmentioning

confidence: 99%

“…Assessment of wound healing by monitoring the cellular volume, dry mass, and refractive index was presented in Ref. 19. The study of cancer cell growth and drug response by monitoring cell dry mass is described in Ref.…”

Section: Introductionmentioning

confidence: 99%

Automated classification of cell morphology by coherence-controlled holographic microscopy

et al. 2017

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“…In addition, the averaged phase contrast ϕ Δ caused by the cells in the area S c and the corresponding average cell thickness d was retrieved. Finally, from the parameters S c and ϕ Δ the change of the cellular dry mass DM, which is related to the intracellular protein content [35], was retrieved as described in [36] following an approach in [37]. Figure 5(b) shows that the area S c covered by the cells is underestimated for the quantitative phase images from single holograms in comparison to the averaged phase images from modulated illumination for periods in which the cells adhere thin on the Petri dish bottom.…”

Section: Resultsmentioning

confidence: 99%

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

Schubert

Vollmer

Ketelhut³

et al. 2014

Biomed. Opt. Express

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Self-interference digital holographic microscopy (DHM) has been found particular suitable for simplified quantitative phase imaging of living cells. However, a main drawback of the self-interference DHM principle are scattering patterns that are induced by the coherent nature of the laser light which affect the resolution for detection of optical path length changes. We present a simple and efficient technique for the reduction of coherent disturbances in quantitative phase images. Therefore, amplitude and phase of the sample illumination are modulated by an electrically focus tunable lens. The proposed method is in particular convenient with the selfinterference DHM concept. Results from the characterization of the method show that a reduction of coherence induced disturbances up to 70 percent can be achieved. Finally, the performance for enhanced quantitative imaging of living cells is demonstrated.

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“…In conclusion, DH microscopy analysis opens a novel diagnostic option for optical quantification of inflammation in murine models of colitis. Moreover, the same research group assessed cellular growth and motility in epithelial wound healing in vitro using DH microscopy [53]. Interestingly, phase images quantifying cell thickness, dry mass, and tissue density were demonstrated.…”

Section: Quantification Of Inflammationmentioning

confidence: 99%

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

El-Schich¹,

Kamlund²,

Janicke³

et al. 2017

Holographic Materials and Optical Systems

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Digital holographic (DH) microscopy is a digital high-resolution holographic imaging technique with the capacity of quantification of cellular conditions without any staining or labeling of cells. The unique measurable parameters are the cell number, cell area, thickness, and volume, which can be coupled to proliferation, migration, cell cycle analysis, viability, and cell death. The technique is cell friendly, fast and simple to use and has unique imaging capabilities for time-lapse investigations on both the single cell and the cell-population levels. The interest for analyzing specifically cell volume changes with DH microscopy, resulting from cytotoxic treatments, drug response, or apoptosis events has recently increased in popularity. We and others have used DH microscopy showing that the technique has the sensitivity to distinguish between different cells and treatments. Recently, DH microscopy has been used for cellular diagnosis in the clinic, providing support for using the concept of DH, e.g., screening of malaria infection of red blood cells (RBC), cervix cancer screening, and sperm quality. Because of its quick and label-free sample handling, DH microscopy will be an important tool in the future for personalized medicine investigations, determining the optimal therapeutic concentration for both different cancer types and individual treatments.

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Quantitative Stain-Free and Continuous Multimodal Monitoring of Wound Healing In Vitro with Digital Holographic Microscopy

Cited by 81 publications

References 61 publications

Automated classification of cell morphology by coherence-controlled holographic microscopy

Automated classification of cell morphology by coherence-controlled holographic microscopy

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

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

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