An image-based flow cytometric approach to the assessment of the nucleus-to-cytoplasm ratio
| Autor: | Elizabeth S. L. Berndl, Michael J. Moore, Michael C. Kolios, Joseph A. Sebastian |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Cytoplasm
Cell Spectrum Analysis Techniques 0302 clinical medicine Neoplasms Breast Tumors Medicine and Health Sciences Image Cytometry Staining 0303 health sciences Multidisciplinary medicine.diagnostic_test Chemistry Cell Staining Flow Cytometry medicine.anatomical_structure Oncology Nephrology Spectrophotometry Renal Cancer 030220 oncology & carcinogenesis Medicine Cell lines Cytophotometry Anatomy Biological cultures Research Article Histology Imaging Techniques Science Flow cytometry 03 medical and health sciences HT29 Cells Malignant Tumors Breast Cancer Fluorescence Imaging medicine Humans HT29 cells 030304 developmental biology Cell Nucleus Cancers and Neoplasms Biology and Life Sciences Molecular biology Research and analysis methods Specimen Preparation and Treatment Cell culture Cancer cell Nucleus |
| Zdroj: | PLoS ONE PLoS ONE, Vol 16, Iss 6, p e0253439 (2021) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0253439 |
| Popis: | 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±1.2μm and 9.0±1.1μm for OCI-AML5 cells, 24.5±2.6μm and 15.6±2.1μm for CAKI-2 cells, 16.2±1.8μm and 11.2±1.3μm for HT29 cells, 18.0±3.7μm and 12.5±2.1μm for SK-BR-3 cells, and 19.4±2.2μm and 10.1± 1.8μm 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. |
| Databáze: | OpenAIRE |
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