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

Abstract

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

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

“…Thus, we gated the fluorescent channel images based on their gradient root-mean-squared values to isolate cell images that were highly focused to distinguish between the nucleus and other parts of the cell (see Fig 1B [32], slightly larger than SK-BR-3 cell sizing using microfluidic cytometry [33], and in good agreement with published values of MCF-10As [34]. Moreover, our previous studies with MCF-7, PC-3 and MDA-MB-231 cells were all in good agreement with other validated techniques [25][26][27]. The nuclear diameters of these cell lines follow a similar trend, with CAKI-2 cells (15.6 ± 2.1 μm) having the largest nuclei, followed by SK-BR-3 cells (12.…”

“…The rationale for this gate is based on the visual clustering of cells we see in the scatter plot based on area and high pixel intensity. Masks used for the image analysis process following the same protocol as our previously published work to accurately measure cell diameter and can be seen in Fig 2B–2D [ 25 – 27 ]. Here, the eroded masks were used to determine the cell diameter where IDEAS provided the diameter of a circle that has the same area as the eroded masks [ 28 ].…”

“…Previously, our group has used IFC to compare cell size measurements with measurements done by photoacoustic microscopy and photoacoustic flow cytometry which were used for the N:C analysis of cultured breast and prostate cancer cells [ 25 – 27 ]. In this work, we determine the N:C ratio of four different malignant cell lines each originating from different tissues and the N:C ratio of a single non-malignant cell line.…”

The dance of the nanobubbles: detecting acoustic backscatter from sub-micron bubbles using ultra-high frequency acoustic microscopy

“…Other bulk particle sizing techniques, such as DLS, are sensitive to these inhomogeneities and report a mixed result describing both bubble and particulate size. In the future, if individual NBs can be isolated and measured individually, for example by flowing a diluted sample of bubbles through an acoustic flow cytometer, 29 the feasibility of applying backscattering models such as those that are used for sizing biological cells [26][27][28] for the purposes of sizing individual NBs can also be investigated.…”

“…18,19 Analysis of the RF-signals recorded in UHF pulse-echo studies allows for extraction of information such as the mechanical properties of biological cells, 20 and mitotic cell cycle phase. 21 Furthermore, when the wavelength of the insonifying acoustic pulse is comparable to the dimensions of the scattering object, unique features in the backscattered power spectrum [22][23][24] enable the sizing of microscopic objects such as spherical microbeads 25 and single cells [26][27][28] without the need for 2D scanning. Such techniques have previously been utilized in UHF acoustic flow cytometry systems for the purposes of sizing cancer cells to a high degree of accuracy and demonstrate good agreement with gold standard particle sizing techniques, such as the Coulter Counter.…”