Abstract

Separating specific cell phenotypes from a heterotypic mixture is a critical step in many research projects. Traditional methods usually require a large sample volume and a complex preparation process that may alter cell property during the sorting process. Here we present the use of electrical impedance as an indicator of cell health and for identifying specific microalgal phenotypes. We developed a microfluidic platform for measuring electrical impedance at different frequencies using the bacterium-sized green alga Picochlorum SE3. The cells were cultured under different salinity conditions and sampled at four different time points. Our results demonstrate the utility of electrical impedance as an indicator of cell phenotype by providing results that are consistent with known changes in cell size and physiology. Outliers in the cell data distribution are particularly useful because they represent phenotypes that have the ability to maintain size and/or membrane ionic permeability under prolonged salt stress. This suggests that our device can be used to identify and sort desired (e.g., experimentally evolved, mutant) cell phenotypes based on their electrical impedance properties.

Highlights

  • Separating specific cell phenotypes from a heterotypic mixture is a critical step in many research projects

  • With the aid of microfluidic flow cytometry technology, impedance spectroscopy requires a smaller sample volume when compared to traditional methods, while maintaining high sensitivity[23,24,25]

  • Strategies for maintaining ionic homeostasis are critical for the survival of Picochlorum SE3 in its natural habitat of a brackish water coastal lagoon that is subject to large fluctuations in salinity through evaporation, precipitation, and tidal influx of seawater

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Summary

Introduction

Separating specific cell phenotypes from a heterotypic mixture is a critical step in many research projects. Outliers in the cell data distribution are useful because they represent phenotypes that have the ability to maintain size and/or membrane ionic permeability under prolonged salt stress This suggests that our device can be used to identify and sort desired (e.g., experimentally evolved, mutant) cell phenotypes based on their electrical impedance properties. To achieve single-cell resolution of gene expression patterns requires the more specialized tools of single cell transcriptomics that may be limited to smaller sample sizes due to the costs of generating individual libraries for 100 s or 1000 s of cells, followed by high-throughput sequencing[4,5] Given these considerations, there is a need to develop tools with single cell resolution that provide meaningful insights into cell health and can be applied to millions of cells at low cost. We demonstrate the utility of electrical impedance as a phenotype indicator that reflects the change in size and permeability of cells under different salt stresses

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