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This version was published on April 1, 2007
Journal of Biomolecular Screening, Vol. 12, No. 3, 361-369 (2007)
DOI: 10.1177/1087057106297565

Turbo-Mixing in Microplates

Elisa Mitre

Institute of Biochemistry, Medical Faculty, Friedrich Schiller University Jena, Jena, Germany

Margarete Schulze

Institute of Biochemistry, Medical Faculty, Friedrich Schiller University Jena, Jena, Germany

Gerhard A. Cumme

Institute of Biochemistry, Medical Faculty, Friedrich Schiller University Jena, Jena, Germany

Fabian Rößler

Institute of Biochemistry, Medical Faculty, Friedrich Schiller University Jena, Jena, Germany

Torsten Rausch

CyBio AG Jena, Jena, Germany

Heidrun Rhode

Institute of Biochemistry, Medical Faculty, Friedrich Schiller University Jena, Jena, Germany, Heidrun.Rhode{at}mti.uni-jena.de

How to effectively mix small volumes of liquids within microplate wells is a still underestimated and often neglected challenge. The method the authors introduce here relies on violent turbulent motion within a liquid caused by spotting an organic solvent drop onto its surface. The amount needed, less than 1 to 3 µL, is generally small enough not to alter bioactive molecules. Moreover, a solvent may be selected for its compatibility with assay components. The method was tested with layers of aqueous liquids that differ in pH and concentration of a pH-dependent dye, allowing mixing to be monitored optically. Rapid mixing was caused by spotting drops of alcohols, acetone, acetonitrile, and aqueous solutions of these, as long as the difference of surface tension between the drop and the uppermost layer of the bulk liquid surpassed 30 dynes/cm. Along with this difference, position and velocity of spotting, as well as viscosity and geometry of the bulk liquid volume, may influence the turbulence evoked. No significant difference was found for the activity of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase when measured after mixing by shaking and after mixing by spotting 1 µL of methanol onto assays within 96-well microplates. (Journal of Biomolecular Screening 2007:361-369)

Key Words: microplate • turbulent mixing • Marangoni convection • thermocapillary convection • µL assay mixing


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