Image Analysis to Study Cytoskeleton Alterations in Cancer Cells Exposed to Nanosecond Pulsed Electric Field

The cytoskeleton is a dynamical protein structure that plays a key role in cellular physiological processes. The assessment of the cytoskeleton dynamics is of prime importance to reveal mechanisms involved in cell adaptation to any type of stress. Fluorescence imaging of cytoskeleton structures enables analysis of the impact of mechanical stimulation on the cytoskeleton, e.g. the exposure to nanosecond pulsed electric field (nsPEF). NsPEF utilizes pulses in nanosecond range (from few up to 300 ns) and high voltage (up to 100 kVs), which can significantly affect all cell structures. This method can be applied in medicine, industry and molecular biology for the efficient molecule transport across the cell membranes. The detailed analysis of the separate cell structures and protein organization seems to be important in the overall evaluation of the nsPEF impact on treated cells. The automatic image analysis seems crucial for proper evaluation of the therapeutic efficiency. The aim of the study is a computational approach for efficient F-actin filament analysis in cells exposed to electroporation process in the nanosecond range. The research was performed on cancer cells derived from gastrointestinal tract exposed to the nanosecond pulsed electric fields in relation to untreated controls. The presented tool might be useful in precise evaluation of the electroporation protocols effectiveness in cancer and normal cells. The proposed methodology showed high sensitivity values and similar accuracy compared to state-of-the-art methods.