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Proceedings Paper

Comparison of measurements of the nucleus-to-cytoplasmic ratio in MCF-7 cells using ultra-high frequency photoacoustic microscopy and imaging flow cytometry
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Paper Abstract

A cell’s nucleus-to-cytoplasm ratio (N:C) can be used as a metric in histology for staging malignancy. Current techniques used for N:C assessment are time-consuming, low-throughput, and lack 3-D structure assessment. In this study, we assess the N:C ratio of MCF-7 cells using an ultra-high frequency acoustic/photoacoustic (PA) microscope and compare measurements to an imaging flow cytometer (IFC). MCF-7 cells were stained with the DRAQ-5 nuclear dye to facilitate production of fluorescence and PA signals. A PA microscope (PAM) consisting of a 375 MHz transducer and a pulsed 532 nm laser focused through a 10X optical objective was used for cell measurements. Cell and nucleus diameters of 37 stationary MCF-7 cells were obtained by fitting the power spectrum of backscattered ultrasound pulses and emitted PA waves, respectively, to well-established theoretical models. 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 2004 MCF-7 cells from the IFC. For both systems, the N:C ratio was calculated as the ratio ofthe nucleus diameter to total cell diameter. The average cell and nucleus diameters from PAM were 15.2 ± 3.5 μm and 10.2 ± 3.5 μm, respectively, and were in good agreement with the average cell and nucleus diameters of 18.7 ± 2.6 μm and 12.6 ± 1.9 μm measured by the IFC. The N:C ratios were in excellent agreement, calculated to be 0.68 ± 0.19 and 0.68 ± 0.09 for PAM and IFC, respectively. This study demonstrates the ability of PAM to measure the N:C ratio of cells which in excellent agreement with IFC assessments.

Paper Details

Date Published: 27 February 2019
PDF: 7 pages
Proc. SPIE 10878, Photons Plus Ultrasound: Imaging and Sensing 2019, 108784M (27 February 2019); doi: 10.1117/12.2507508
Show Author Affiliations
Joseph A. Sebastian, Ryerson Univ. (Canada)
Institute for Biomedical Engineering, Science and Technology, St. Michael's Hospital (Canada)
Keenan Research Ctr. for Biomedical Science, St. Michael's Hospital (Canada)
Michael J. Moore, Ryerson Univ. (Canada)
Institute for Biomedical Engineering, Science and Technology, St. Michael's Hospital (Canada)
Keenan Research Ctr. for Biomedical Science, St. Michael's Hospital (Canada)
Michael C. Kolios, Ryerson Univ. (Canada)
Institute for Biomedical Engineering, Science and Technology, St. Michael's Hospital (Canada)
Keenan Research Ctr. for Biomedical Science, St. Michael's Hospital (Canada)


Published in SPIE Proceedings Vol. 10878:
Photons Plus Ultrasound: Imaging and Sensing 2019
Alexander A. Oraevsky; Lihong V. Wang, Editor(s)

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