An impedimetric assay for the identification of abnormal mitochondrial dynamics in living cells

Abstract

Mitochondrial dynamics (fission and fusion) plays an important role in cell functions. Disruption in mitochondrial dynamics has been associated with diseases such as neurobiological disorders and cardiovascular diseases. Analysis of mitochondrial fission/fusion has been mostly achieved through direct visualization of the fission/fusion events in live-cell imaging of fluorescently labeled mitochondria. In this study, we demonstrated a labelfree, non-invasive Electrical Impedance Spectroscopy (EIS) approach to analyze mitochondrial dynamics in a genetically modified human neuroblastoma SH-SY5Y cell line with no huntingtin protein expression. Huntingtin protein has been shown to regulate mitochondria dynamics. We performed EIS studies on normal SH-SY5Y cells and two independent clones of huntingtin-null cells. The impedance data was used to determine the suspension conductivity and further cytoplasmic conductivity and relate to the abnormal mitochondrial dynamics. For instance, the cytoplasm conductivity value was increased by 11% from huntingtin-null cells to normal cells. Results of this study demonstrated that EIS is sensitive to characterize the abnormal mitochondrial dynamics that can be difficult to quantify by the conventional microscopic method.

The Use of Tissue Engineering to Fabricate Perfusable 3D Brain Microvessels in vitro

“…In corporation of analytic measurements is an area for future improvement. For instance, microfluidic based electrical impedance techniques have been widely used to characterize the barrier related parameters and cellular and physical properties of cells ( Cucullo et al, 2008 ; Douville et al, 2010 ; Szulcek et al, 2014 ; Reiss and Wegener, 2015 ; Bischoff et al, 2016 ; Galpayage Dona et al, 2017 , 2020b ), but few 3D models have real-time continuous measurements ( Cucullo et al, 2007 ). Instead most rely on electrodes inserted into ports connected to the inner and outer luminal compartment requiring cessation of perfusion to obtained single timepoint measurements.…”