Dual-frequency impedance assays for intracellular components in microalgal cells. Issue 3 (24th January 2022)

• Record Type: Journal Article
• Authors: Tang, Tao; Liu, Xun; Yuan, Yapeng; Kiya, Ryota; Shen, Yigang; Zhang, Tianlong; Suzuki, Kengo; Tanaka, Yo; Li, Ming; Hosokawa, Yoichiroh; Yalikun, Yaxiaer
• Is Part Of: Lab on a chip. Volume 22:Issue 3(2022)
• Page Start: 550
• Page End: 559
• Publication Date: 2022-01-24
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/d1lc00721a
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : The capabilities of impedance cytometry to detect intracellular components ( i.e., volume and distribution) are investigated, and it is found that non-uniform intracellular distribution causes the tilt tendency of high-frequency impedance pulses. Abstract : Intracellular components (including organelles and biomolecules) at the submicron level are typically analyzed in situ by special preparation or expensive setups. Here, a label-free and cost-effective approach of screening microalgal single-cells at a subcellular resolution is available based on impedance cytometry. To the best of our knowledge, it is the first time that the relationships between impedance signals and submicron intracellular organelles and biomolecules are shown. Experiments were performed on Euglena gracilis ( E. gracilis ) cells incubated under different incubation conditions ( i.e., aerobic and anaerobic) and 15 μm polystyrene beads (reference) at two distinct stimulation frequencies ( i.e., 500 kHz and 6 MHz). Based on the impedance detection of tens of thousands of samples at a throughput of about 900 cells per second, three metrics were used to track the changes in biophysical properties of samples. As a result, the electrical diameters of cells showed a clear shrinkage in cell volume and intracellular components, as observed under a microscope. The morphology metric of impedance pulses ( i.e., tilt index) successfully characterized the changes in cell shape and intracellular composition distribution. Besides, the electrical opacity showed a stable ratio of the intracellular components to cell volume under the cellular self-regulation. Additionally, simulations were used to support these findings and to elucidate how submicron intracellular components and cell morphology affect impedance signals, providing a basis for future improvements. This work opens up a label-free and high-throughput way to analyze single-cell intracellular components by impedance cytometry.