Mitigation of Volatile Chemicals' Effect on Adjacent Microtiter Plate Wells

Abstract

In vitro new approach methodologies used to assess chemicals for biological effects are typically designed to limit the amount of test article required and to promote efficiencies such as compatibility with liquid handlers, and so forth. This is certainly true in the case of genetic toxicology, where many methods have been and continue to be developed with 96- or 384-well plate processing in mind. However, one recognized concern with microwell plates is that the volatility of test substances and/or their metabolites and/or their degradation products may affect adjacent wells. Here, we describe an approach that combines breathable membranes as well as activated carbon filters to mitigate volatility issues in 96 well plates. These experiments were performed with cyclophosphamide- and trimethylhydroquinone-exposed TK6 cells and utilized the biomarkers micronuclei, p53, γH2AX, phospho-histone H3, and nuclei to counting bead ratios to both demonstrate volatility impact and to assess the effectiveness of the solution described herein.