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

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 maligna...

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Autores principales:	Joseph A Sebastian, Michael J Moore, Elizabeth S L Berndl, Michael C Kolios
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d8dbb4f2bb3f436490ec0dbe97595209
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spelling	oai:doaj.org-article:d8dbb4f2bb3f436490ec0dbe975952092021-12-02T20:10:08ZAn image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.1932-620310.1371/journal.pone.0253439https://doaj.org/article/d8dbb4f2bb3f436490ec0dbe975952092021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0253439https://doaj.org/toc/1932-6203The 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.Joseph A SebastianMichael J MooreElizabeth S L BerndlMichael C KoliosPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 6, p e0253439 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Joseph A Sebastian Michael J Moore Elizabeth S L Berndl Michael C Kolios An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
description	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.
format	article
author	Joseph A Sebastian Michael J Moore Elizabeth S L Berndl Michael C Kolios
author_facet	Joseph A Sebastian Michael J Moore Elizabeth S L Berndl Michael C Kolios
author_sort	Joseph A Sebastian
title	An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
title_short	An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
title_full	An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
title_fullStr	An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
title_full_unstemmed	An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
title_sort	image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/d8dbb4f2bb3f436490ec0dbe97595209
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An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio.

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