Microfluidic Cytometry for High-Throughput Characterization of Single Cell Cytoplasmic Viscosity Using Crossing...

This article presents an approach of microfluidic flow cytometry capable of continuously characterizing cytoplasmic viscosities of single cells. The microfluidic system consists of a major constriction channel and a side constriction channel perpendicularly crossing each other. Cells are forced to rapidly travel through the major channel and are partially aspirated into the side channel when passing the channel junction. Numerical simulations were conducted to model the time dependence of the aspiration length into the side channel, which enables the measurement of cytoplasmic viscosity by fitting the model results to experimental data. As a demonstration for high-throughput measurement, the cytoplasmic viscosities of HL-60 cells that were native or treated by N-Formylmethionine-leucyl-phenylalanine (fMLP) were quantified with sample sizes as large as thousands of cells. Both the average and median cytoplasmic viscosities of native HL-60 cells were found to be about 10% smaller than those of fMLP-treated HL-60 cells, consistent with previous observations that fMLP treatment can increase the rigidity of white blood cells. Furthermore, the microfluidic system was used to process granulocytes from three donors (sample size >1,000 cells for each donor). The results revealed that the cytoplasmic viscosity of granulocytes from one donor was significantly higher than the other two, which may result from the fact that this donor just recovered from an inflammation. In summary, the developed microfluidic system can collect cytoplasmic viscosities from thousands of cells and may function as an enabling tool in the field of single-cell analysis. © 2019 International Society for Advancement of Cytometry.