an-electroporation-cytometry-protocol-for-live-cell-fluorescence

An Electroporation Cytometry Protocol for Live-Cell, Fluorescence Microscopy Using U2 OS Cell Culture

Pulsed electric fields (PEFs) have a wide range of applications in medical research and clinical applications. A key area of research focuses on ...

Toronto Metropolitan University

Science and Technology (iBEST), a partnership between St. Michael's Hospital, a site of Unity Health Toronto, and Toronto Metropolitan University

Keenan Research Centre for Biomedical Science at St. Michael's Hospital

Electric fields are used in biology to address a broad range of questions and through a variety of techniques, including electroporation, gene electrotransfer (GET), electrostimulation (ES), and electrochemotherapy. Each of these modalities requires specific conditions and has drastically different target outcomes on the cell. ES has demonstrated that non-pore forming electric fields alter cell cycle progression. However, pore forming electric fields such as with GET have not been as widely explored despite major clinical advancements. Additionally, the real-time visual analysis of electrical field effects on mammalian cell culture is currently lacking among most commercial systems. To facilitate investigations into these research areas, an electroporation cytometry system was developed including a custom chamber compatible with live cell imaging and exponential decay pulse generator for live cell analysis. The functionality of the system was demonstrated using a recombinant cell line using U-2 OS cells and FUCCI(CA)5 cell cycle reporter. The exposure of the cells to a 180 V pulse in both unsynchronized and synchronized populations revealed an effect on the cell cycle.

Thomas Nesmith

Biography

JoVE Journal Publications

Other Publications