Incubator-independent cell-culture perfusion platform for continuous long-term microelectrode array electrophysiology and time-lapse imaging

Dirk Saalfrank, Anil Krishna Konduri, Shahrzad Latifi, Rouhollah Habibey, Asiyeh Golabchi, Aurel Vasile Martiniuc, Alois Knoll, Sven Ingebrandt, Axel Blau

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Most in vitro electrophysiology studies extract information and draw conclusions from representative, temporally limited snapshot experiments. This approach bears the risk of missing decisive moments that may make a difference in our understanding of physiological events. This feasibility study presents a simple benchtop cell-culture perfusion system adapted to commercial microelectrode arrays (MEAs), multichannel electrophysiology equipment and common inverted microscopy stages for simultaneous and uninterrupted extracellular electrophysiology and time-lapse imaging at ambient CO2 levels. The concept relies on a transparent, replica-casted polydimethylsiloxane perfusion cap, gravity- or syringe-pump-driven perfusion and preconditioning of pH-buffered serum-free cellculture medium to ambient CO2 levels at physiological temperatures. The low-cost microfluidic in vitro enabling platform, which allows us to image cultures immediately after cell plating, is easy to reproduce and is adaptable to the geometries of different cell-culture containers. It permits the continuous and simultaneousmultimodal long-term acquisition or manipulation of optical and electrophysiological parameter sets, thereby considerably widening the range of experimental possibilities. Two exemplary proofof- concept long-term MEA studies on hippocampal networks illustrate system performance. Continuous extracellular recordings over a period of up to 70 days revealed details on both sudden and gradual neural activity changes in maturing cell ensembles with large intra-day fluctuations. Correlated time-lapse imaging unveiled rather static macroscopic network architectures with previously unreported local morphological oscillations on the timescale of minutes.

Original languageEnglish
Article number150031
JournalRoyal Society Open Science
Volume2
Issue number6
DOIs
StatePublished - Jun 2015

Keywords

  • Benchtop perfusion
  • Cell culture
  • MEA electrophysiology
  • Polydimethylsiloxane
  • Replica casting
  • Time-lapse imaging

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