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Particle Self-Aligning, Focusing, and Electric Impedance Microcytometer Device for Label-Free Single Cell Morphology Discrimination and Yeast Budding Analysis
journal contribution
posted on 2019-10-18, 12:34 authored by Xinwu Xie, Zhiwei Zhang, Xiang Ge, Xiaohao Zhao, Limei Hao, Zhen Cheng, Weibin Zhou, Yaohua Du, Lei Wang, Feng Tian, Xinxi XuMicrofluidic
electric impedance flow cytometry (IFC) devices have
been applied in single cell analysis, such as cell counting, volume
discrimination, cell viability, etc. A cell’s shape provides
specific information about cellular physiological and pathological
conditions, especially in microorganisms such as yeast. In this study,
the particle orientation focusing was theoretically analyzed and realized
by hydrodynamics. The pulse width (passing time for the particles)
of the conductance signal was used to discriminate particle shapes.
Spherical and rod-shaped particles with similar volumes/lengths were
differentiated by the IFC device, using the impedance pulse parameters
of the events. Then, typical late-budding, early budding, and unbudded
yeast cells were distinguished by the width, amplitude, and ratio
of width to amplitude (R) of the impedance pulse.
The pulse amplitude and the R combination gate for
identifying the late-budding yeast was estimated through the statistic
results. Using the gate, the late-budding rates under different conditions
were calculated. Late-budding rates obtained using our method showed
a high correlation (R2 = 0.83) with the
manual cell counting result and represented the budding status of
yeast cells under different conditions proficiently. Thus, the late-budding
rate calculated using the above method can be used as a qualitative
parameter to assess the reproductive performance of yeast and whether
a yeast culturing environment is optimal. This IFC device and cell
shape discrimination method is very simple and could be applied in
the fermentation industry and other microorganisms’ discrimination
as a rapid analysis technique in the future.
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Keywords
Label-Free Single Cell Morphology DiscriminationIFC devicelate-buddingYeast Budding Analysis Microfluidicimpedance pulse parametersunbudded yeast cellswidthElectric Impedance Microcytometer Devicecell shape discrimination methodimpedance flow cytometryR combination gateamplitudeyeast culturing environment
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