Label‐free biomarkers of human embryonic stem cell differentiation to hepatocytes

Tsikritsis, Dimitrios; Shi, Hang; Wang, Yuan; Velugotla, Srinivas; Sršeň, Vlastimil; Elfick, Alistair; Downes, Andrew

doi:10.1002/cyto.a.22875

Cited by 15 publications

(14 citation statements)

References 70 publications

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“…Hepatoma cells with different metastasis ability showed disparity in terms of the distribution patterns of Young’s modulus ( Tian et al, 2015 ). Differences in elastic modulus are found between normal liver, hepatoma, liver embryonic stem cells ( Kim et al, 2013 ; Sun et al, 2016 ; Tsikritsis et al, 2016 ). An AFM topology study showed that the increased surface roughness and reduced cytoskeleton height are associated with curcumin-induced G2/M phase arrest in HepG2 cells ( Jiang et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

Effects of Lipid Deposition on Viscoelastic Response in Human Hepatic Cell Line HepG2

Yang

et al. 2021

Front. Physiol.

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Hepatic steatosis is associated with various liver diseases. The main pathological feature of steatosis is the excessive lipid accumulation. Ultrasound has been extensively used for the diagnosis of hepatic steatosis. However, most ultrasound-based non-invasive methods are still not accurate enough for cases with light lipid infiltration. One important reason is that the extent to which lipid infiltration may affect mechanical properties of hepatocytes remains unknown. In this work, we used atomic force microscope and in vitro dose-dependent lipid deposition model to detect the quantitative changes of mechanical properties under different degrees of steatosis in a single-cell level. The results show that hepatic cells with lipid deposition can be treated as linear viscoelastic materials with the power law creep compliance and relaxation modulus. Further analysis showed that even slight accumulation of lipid can lead to measurable decrease of stiffness and increased fluidity in liver cells. The accurate detection of viscoelastic properties of hepatocytes and the analysis methods may provide novel insights into hepatic steatosis grading, especially in the very early stage with reversible liver lesion. The application of viscoelasticity index for grading fat deposition might be a new detection indicator in future clinical diagnosis.

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Section: Introductionmentioning

confidence: 99%

Effects of Lipid Deposition on Viscoelastic Response in Human Hepatic Cell Line HepG2

Yang

et al. 2021

Front. Physiol.

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“…There is also research into using DEP to concentrate relevant biomarkers for chemical assay . Much of the published efforts into electrokinetic separations for diagnostics have been focused in cell sorting, e.g., .…”

Section: Introductionmentioning

confidence: 99%

Electrokinetic device design and constraints for use in high conductance solutions

2017

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The quest for new cell-free DNA and exosome biomarker-based molecular diagnostics require fast and efficient sample preparation techniques. Conventional methods for isolating these biomarkers from blood are both time-consuming and laborious. New electrokinetic microarray devices using dielectrophoresis (DEP) to isolate cell-free DNA and exosome biomarkers have now greatly improved the sample preparation process. Nevertheless, these devices still have some limitations when used with high conductance biological fluids, e.g. blood, plasma, and serum. This study demonstrates that electrochemical damage may occur on the platinum electrodes of DEP microarray devices. It further examines two model device designs that include a parallel wire arrangement and a planar array. Effective isolation of fluorescent beads with parallel wires is shown under low-conductance conditions (10 S/m), but electrothermal flow overcomes DEP forces under high conductance conditions (>0.1 S/m). Planar devices are shown to be effective under high conductance conditions (∼1 S/m) without the deleterious effects of electrothermal flow. This study provides new insights into design compromises and limitations for producing future electrokinetic devices for better performance with high conductance solutions.

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“…Threedimensional quantitative phase images of cells are reconstructed from several hundred frames, and the volume, dry mass, and density recorded. Still, the labelfree nature of quantitative phase signatures is attractive for non-destructive cell sorting, and may be added to the list of list of label-free techniques useful for such purposes, including 2D light scattering (6), Raman spectroscopy, stimulated Raman scattering, dielectrophoresis, and atomic force microscopy, to name a few (7). Projections compared to dense multi-frame acquisition would seem to favor higher throughput.…”

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confidence: 99%

“…These two studies highlight the tradeoff between acquisition throughput versus phase and spatial resolution/information density, which is balanced in the study by Roitshtain, Shaked and coauthors (2). Still, the labelfree nature of quantitative phase signatures is attractive for non-destructive cell sorting, and may be added to the list of list of label-free techniques useful for such purposes, including 2D light scattering (6), Raman spectroscopy, stimulated Raman scattering, dielectrophoresis, and atomic force microscopy, to name a few (7). While these techniques are sensitive to different properties of cells-phase signatures, Mie scattering, biochemistry, lipid droplets, capacitance, and elasticity, in order, for the techniques listed above-they each allow cells to retain viability for future therapeutic or functional testing.…”

mentioning

confidence: 99%

Holography, machine learning, and cancer cells

Raub

Nehmetallah

2017

Cytometry Pt A

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COMMENTARY LIQUID biopsy to detect circulating solid tumor cells, cellfree DNA or tumor exosomes in peripheral blood holds great promise as a minimally invasive method to detect cancer at an early stage, plan treatments, and monitor disease response to therapy (1). Two great challenges to detecting cancer cells in whole blood are the low relative abundance of tumor cells in proportion to erythrocytes and leukocytes, and the need to introduce highly-specific exogenous labels to identify tumor biomarkers. A recently published article in Cytometry Part A (2) detects cancer cells using a rapid, label-free technique based on machine learning from quantitative phase/digital holographic microscopy datasets. The results demonstrate automated discrimination of normal and cancer cell types flowing in a microfluidic channel, pointing to the potential for high-throughput processing of circulating tumor cells in an imaging flow cytometer. The method is compatible with microfluidic approaches to sequester or enrich circulating tumor cells from normal blood cells and whole blood, respectively.Without staining, biological cells are transparent under bright field microscopy resulting in low contrast intensity images. In that article, Shaked and coauthors (2) utilize an internal contrast mechanism based on the index of refraction. Contrary to traditional qualitative phase contrast (PC) techniques, such as Zernike's PC and differential interference contrast microscopy, this off-axis interferometric label-free approach can distinguish normal cells from cancer cell lines of different disease stages using quantitative textural and morphological parameters derived from the segmented optical thickness or optical path delay (OPD) of the cell on all spatial points. This OPD is the integral of the index of refraction at each pixel of a projection across the cells' thickness. To use OPD parameters for sensitive cell screening, the phase signal must possess high fidelity and consistency across images and during the imaging session.The optical setup used in this study is a low-coherence off-axis interferometer with a Mach-Zehnder configuration. The source used is a supercontinuum laser coupled to an acousto-optical tunable filter with a 7 nm bandwidth. The low-coherence source is used to reduce speckle noise. Retroreflectors are used to adjust the sample and reference beam paths to enhance fringe visibility. Stationary aberrations and field curvature aberrations are compensated for using empty sample measurements. Phase unwrapping was performed using a standard unweighted least square algorithm to obtain the OPD maps of the cells. A normalized cut-edge detector algorithm, which is based on graph formulation, was used to separate cells from their background. This algorithm was followed by morphological image operations (opening, dilation, and thresholding) to connect the gradient lines and to remove background pixels erroneously detected. The cells from cultured cell lines, flowed through a microfluidic channel and imaged at high frame rates, ...

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Label‐free biomarkers of human embryonic stem cell differentiation to hepatocytes

Cited by 15 publications

References 70 publications

Effects of Lipid Deposition on Viscoelastic Response in Human Hepatic Cell Line HepG2

Effects of Lipid Deposition on Viscoelastic Response in Human Hepatic Cell Line HepG2

Electrokinetic device design and constraints for use in high conductance solutions

Holography, machine learning, and cancer cells

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