Comparison of two- and three-dimensional nuclear to cytoplasm ratios in analyzing lung cancer cell lines

Huang, Hsu-Cheng; Chiang, Shu‐Jen; Wen, Shu-Han; Lee, Pei-Jung; Chen, Huei‐Wen; Chen, Yang‐Fang; Dong, Chongying

doi:10.1117/12.2507400

Cited by 5 publications

(13 citation statements)

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“…[4][5][6] These shortcomings have led to the development of alternative systems for N:C ratio quantification. Emerging technologies such as digital image analysis using HALO imaging-analysis software in urine cytology, 7 immunofluorescence microscopy paired with ImageJ, 8 and 3-D multiphoton microscopy 9 have been used to assess the N:C ratio but all require a lengthy imaging/analysis time for individual cells. Flow cytometry (FC) overcomes the drawback of slow analysis speeds at the expense of direct measurements of cell morphology.…”

Section: Introductionmentioning

confidence: 99%

Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study

2019

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Section: Introductionmentioning

confidence: 99%

Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study

2019

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“…Our N:C results are consistent with international standards of cytopathology (Hang et al's [42] finding of an N:C ratio cutoff value of 0.5 for atypical urothelial cells, and McIntire et al's [43] finding of a N:C ratio cutoff value below 0.7 for high-grade urothelial carcinoma). Previous work by Rahmadwati et al [13] and Huang et al [14,15] have shown that non-malignant cell types have N:C ratios between 0.2-0.4. Given the standard deviations of our non-malignant cell N:C ratios, our measurements fall within this range.…”

Section: Discussionmentioning

confidence: 96%

“…The drawbacks of histology to assess the N:C ratio and reported inaccuracies and inconsistencies in N:C assessments by morphologists and clinicians [9][10][11] have motivated the implementation of new techniques to determine the N:C ratio, including the use of computer vision [13], multi-photon microscopy [14][15][16], Cell-CT [17] and immunohistochemistry analysis techniques [18]. Rahmadwati et al [13] applied k-means clustering to segment nuclei and cytoplasm from background and connective tissue to detect features indicative of normal tissue, pre-cancerous tissue, and malignant tissues in cervical cancer histology images to assess the N:C ratio.…”

Section: Discussionmentioning

confidence: 99%

“…The technique is heavily ROI dependent and details regarding sample size and number of histological images used are not provided. Multi-photon microscopy techniques studied by Huang et al [14,15] suffered from low sample populations (n < 25) and thus, were not representative of the entire cell population and ineffective in high-throughput settings. A two-photon microscopy (TPM) technique was implemented by Su Lim et al [16] to assess the nuclear area and N:C ratios in human colon tissues.…”

Section: Discussionmentioning

confidence: 99%

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An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio

et al. 2021

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The nucleus-to-cytoplasm ratio (N:C) can be used as one metric in histology for grading certain types of tumor malignancy. Current N:C assessment techniques are time-consuming and low throughput. Thus, in high-throughput clinical contexts, there is a need for a technique that can assess cell malignancy rapidly. In this study, we assess the N:C ratio of four different malignant cell lines (OCI-AML-5—blood cancer, CAKI-2—kidney cancer, HT-29—colon cancer, SK-BR-3—breast cancer) and a non-malignant cell line (MCF-10A –breast epithelium) using an imaging flow cytometer (IFC). Cells were stained with the DRAQ-5 nuclear dye to stain the cell nucleus. An Amnis ImageStreamX® IFC acquired brightfield/fluorescence images of cells and their nuclei, respectively. Masking and gating techniques were used to obtain the cell and nucleus diameters for 5284 OCI-AML-5 cells, 1096 CAKI-2 cells, 6302 HT-29 cells, 3159 SK-BR-3 cells, and 1109 MCF-10A cells. The N:C ratio was calculated as the ratio of the nucleus diameter to the total cell diameter. The average cell and nucleus diameters from IFC were 12.3 ± 1.2 μm and 9.0 ± 1.1 μm for OCI-AML5 cells, 24.5 ± 2.6 μm and 15.6 ± 2.1 μm for CAKI-2 cells, 16.2 ± 1.8 μm and 11.2 ± 1.3 μm for HT-29 cells, 18.0 ± 3.7 μm and 12.5 ± 2.1 μm for SK-BR-3 cells, and 19.4 ± 2.2 μm and 10.1 ± 1.8 μm for MCF-10A cells. Here we show a general N:C ratio of ~0.6–0.7 across varying malignant cell lines and a N:C ratio of ~0.5 for a non-malignant cell line. This study demonstrates the use of IFC to assess the N:C ratio of cancerous and non-cancerous cells, and the promise of its use in clinically relevant high-throughput detection scenarios to supplement current workflows used for cancer cell grading.

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Section: Introductionmentioning

confidence: 99%

Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study

Moore¹,

Sebastian²,

Kolios³

2022

Preprint

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<p>While the nucleus-to-cytoplasmic (N:C) ratio has traditionally been used for assessing cell malignancy, most N:C measurement techniques are time-consuming and performed on thin histological sections, which prohibit assessment of three-dimensional cell structure. A combined ultrahigh frequency ultrasound (US) and photoacoustic (PA) technique was used to assess the size and N:C ratio of cultured cancer cells in three dimensions (3D). The diameters of the cells and their stained nuclei were obtained by fitting the power spectrum of backscattered US pulses and emitted PA waves, respectively, to well-established theoretical models. For comparison, an imaging flow cytometer (IFC) was also used to determine the two-dimensional cell and nucleus sizes from large cell populations using brightfield and fluorescence images, respectively. An N:C ratio was calculated for each cell using the quotient of the measured nucleus diameter and the total cell diameter. The mean N:C ratios calculated using the sound-based approach were 0.68, 0.66, and 0.54 for MCF-7, PC-3, and MDA-MB-231 cells, respectively, and were in good agreement with the corresponding values of 0.68, 0.67, and 0.68 obtained using the IFC. The combined US and PA technique, which assesses cellular N:C ratio in 3D, has potential applications in the detection of circulating tumor cells in liquid biopsies. </p>

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Comparison of two- and three-dimensional nuclear to cytoplasm ratios in analyzing lung cancer cell lines

Cited by 5 publications

References 10 publications

Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study

Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study

An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio

Determination of cell nucleus-to-cytoplasmic ratio using imaging flow cytometry and a combined ultrasound and photoacoustic technique: a comparison study

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