Effective stabilization of ethanol levels in multiple-well tissue culture plates

P. Borgs, D. L. Way, M. H. Witte, C. L. Witte

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Underestimation of ethanol (EtOH) volatility in vitro is a potential source of experimental error. EtOH (0-5% in culture medium) was added to 24- or 96-well tissue culture plates with standard low evaporation lids and incubated at 37°C in humidified 7.5% CO2 and 92.5% air. After 72 hours, approximately 70% of the initial EtOH had disappeared from the aqueous phase of the plate (EtOH volatilization). EtOH concentrations gradually decreased in high-concentration wells (1-5%) and increased in low-concentration wells (0-0.1%) over time. This temporal redistribution of EtOH (EtOH diffusion) was detected after only 1 hour of incubation. Parafilm, Blenderm surgical tape, and ELISA plate-sealing tape barriers inconsistently or inadequately prevented EtOH volatilization and diffusion, but a newly designed plate-sealing clamp (PSC) apparatus inhibited this phenomenon. Rat hepatic sinusoidal endothelial cells cultured with the PSC apparatus maintained intact cell membranes for 72 hours and stable levels of monolayer permeability for at least 48 hours. By stabilizing in vitro EtOH concentrations, the PSC apparatus eliminates a potential source of major experimental error.

Original languageEnglish (US)
Pages (from-to)31-35
Number of pages5
JournalAlcohol
Volume10
Issue number1
DOIs
StatePublished - Jan 1 1993

Keywords

  • Endothelial cells
  • Ethanol volatility
  • Evaporation barriers
  • In vitro alcohol
  • Lactate dehydrogenase
  • Monolayer permeability
  • Multiple-well plates
  • Plate-sealing clamp apparatus

ASJC Scopus subject areas

  • Health(social science)
  • Biochemistry
  • Toxicology
  • Neurology
  • Behavioral Neuroscience

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