An electroporation cytometry system for long-term, live cell cycle analysis

Author:

Nesmith Thomas123ORCID,Vieira Christian1ORCID,Rackus Darius G.123ORCID,Gupta Gagan D.1ORCID

Affiliation:

1. Department of Chemistry and Biology, Toronto Metropolitan University 1 , Toronto, Ontario M5B 2K3, Canada

2. Institute for Biomedical Engineering, Science and Technology (iBEST), a partnership between St. Michael's Hospital, Unity Health Toronto and Toronto Metropolitan University 2 , Toronto, Ontario M5B 1W8, Canada

3. Keenan Research Centre for Biomedical Science, St. Michael's Hospital 3 , Toronto, Ontario M5B 1W8, Canada

Abstract

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.

Funder

Natural Sciences and Engineering Research Council of Canada

Toronto Metropolitan University

Publisher

AIP Publishing

Reference24 articles.

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