Digital Holographic Microscopy: A Quantitative Label-Free Microscopy Technique for Phenotypic Screening

Rappaz, Benjamin; Breton, Billy; Shaffer, Etienne; Turcatti, Gerardo

doi:10.2174/13862073113166660062

Cited by 92 publications

(71 citation statements)

References 31 publications

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“…We note that the matrix inversion in (30) is welldefined, even in the absence of regularization. However, in cell imaging applications, it is often desirable that the specimens be well-isolated from the background [12]. Therefore, we combine our data-fidelity term with a TV functional, which is known to preserve discontinuities better than Tikhonov-type regularizations [34].…”

Section: B Discrete Formulationmentioning

confidence: 99%

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Variational Phase Imaging Using the Transport-of-Intensity Equation

Bostan

Froustey

Nilchian

et al. 2016

IEEE Trans. on Image Process.

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Abstract-We introduce a variational phase retrieval algorithm for the imaging of transparent objects. Our formalism is based on the transport-of-intensity equation (TIE), which relates the phase of an optical field to the variation of its intensity along the direction of propagation. TIE practically requires one to record a set of defocus images to measure the variation of intensity. We first investigate the effect of the defocus distance on the retrieved phase map. Based on our analysis, we propose a weighted phase reconstruction algorithm yielding a phase map that minimizes a convex functional. The method is nonlinear and combines different ranges of spatial frequencies-depending on the defocus value of the measurements-in a regularized fashion. The minimization task is solved iteratively via the alternatingdirection method of multipliers. Our simulations outperform commonly used linear and nonlinear TIE solvers. We also illustrate and validate our method on real microscopy data of HeLa cells.

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Section: B Discrete Formulationmentioning

confidence: 99%

“…This allows for the segmentation of cells and other specific structures. The downside of DHM is that it is a hardware-based solution of higher cost compared to bright-field, DIC, and PC microscopies [12]. In this paper, we consider a low-cost label-free phase imaging approach that uses the transport-of-intensity equation (TIE) [13].…”

Section: Introductionmentioning

confidence: 99%

Variational Phase Imaging Using the Transport-of-Intensity Equation

Bostan

Froustey

Nilchian

et al. 2016

IEEE Trans. on Image Process.

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“…In many fields of biology, such as cancer research, 1 drug discovery, 2,3 cell death, 4 phenotypic screening, 5 study of pathological processes, 6,7 or interactions of cells with biomaterials, 8 the microscopy study of cell morphology belongs to essential research methods. Manual observation and evaluation of cell morphology in microscopy images require a trained biologist who performs inspection on every image.…”

Section: Introductionmentioning

confidence: 99%

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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“…The culture vessel will be placed on the microscope for imaging and then replaced in the 37°C incubator, or placed on a heating plate to retain 37°C, during the analysis. Since the first studies on living cells, DH microscopy has been used to study a wide range of different cell types, e.g., protozoa, bacteria, and plant cells, mammalian cells such as nerve cells, stem cells, various tumor cells, bacterial-cell interactions, red blood cells (RBC), and sperm cells [11][12][13][14][15].…”

Section: Introductionmentioning

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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Digital Holographic Microscopy: A Quantitative Label-Free Microscopy Technique for Phenotypic Screening

Cited by 92 publications

References 31 publications

Variational Phase Imaging Using the Transport-of-Intensity Equation

Variational Phase Imaging Using the Transport-of-Intensity Equation

Automated classification of cell morphology by coherence-controlled holographic microscopy

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

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