Measuring the nucleus-to-cytoplasmic ratio in PC-3 cells using photoacoustic flow cytometry and imaging flow cytometry

Joseph A. Sebastian; Vaskar Gnyawali; Lianne Ho Yan So; Michael J. Moore; Scott S. H. Tsai; Michael C. Kolios

doi:10.1117/12.2506434

27 February 2019 Measuring the nucleus-to-cytoplasmic ratio in PC-3 cells using photoacoustic flow cytometry and imaging flow cytometry

Joseph A. Sebastian, Vaskar Gnyawali, Lianne Ho Yan So, Michael J. Moore, Scott S. H. Tsai, Michael C. Kolios

Author Affiliations +

Proceedings Volume 10878, Photons Plus Ultrasound: Imaging and Sensing 2019; 108782H (2019) https://doi.org/10.1117/12.2506434
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

A cell’s nucleus-to-cytoplasm (N:C) ratio is a histological metric used to stage malignant disease. Current N:C assessment methods, such as optical microscopy, are time-consuming, subjective, and low-throughput. Here, we compare the N:C ratios of prostate cancer (PC-3) cells measured by a novel microfluidic PhotoAcoustic Flow Cytometer (PAFC) to those obtained using an Imaging Flow Cytometer (IFC). PC-3 cells were stained with DRAQ-5 nuclear dye and divided into populations measured using the PAFC and IFC. The PAFC consisted of a microfluidic device integrated with a singleelement ultrasound transducer (375 MHz central frequency) and a sub-nanosecond pulsed laser (532 nm). Individual cells were 3D flow-focused through the overlapping focal region of the ultrasound and laser pulses. PAFC estimation of the cell and nucleus diameters were determined through power spectra fitting of backscattered US waves and emitted PA waves to established theoretical models. An ImageStreamX® IFC was used to acquire brightfield and fluorescent images of individual cells, which were masked, gated, and used to assess the cell (brightfield) and nucleus (fluorescence) diameter to validate the PAFC measurements. The average cell and nucleus diameters determined using the PAFC (n = 388) were 18.8 ± 3.3 μm and 14.3 ± 2.9 μm, respectively. The corresponding values from the IFC (n = 4651) were 18.3 ± 2.2 μm and 12.2 ± 1.9 μm. The N:C ratio (calculated as the ratio of the nucleus diameter to cell diameter) was 0.77 ± 0.10 using the PAFC and 0.67 ± 0.07 using the IFC. Our novel PAFC device has the potential to be used for circulation tumor cell detection using the N:C ratios of cells.

Conference Presentation

Citation Download Citation

Joseph A. Sebastian, Vaskar Gnyawali, Lianne Ho Yan So, Michael J. Moore, Scott S. H. Tsai, and Michael C. Kolios "Measuring the nucleus-to-cytoplasmic ratio in PC-3 cells using photoacoustic flow cytometry and imaging flow cytometry", Proc. SPIE 10878, Photons Plus Ultrasound: Imaging and Sensing 2019, 108782H (27 February 2019); https://doi.org/10.1117/12.2506434

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available