Miniaturization of Mitotic Index Cell-Based Assay Using “Wall-Less” Plate Technology

Abstract

The use of microscopic imaging for the accurate assessment of cells in mitosis is hampered by the round morphology of mitotic cells, which renders them poorly adherent and highly susceptible to loss during the washing stage of cell-based assays. Here, to circumvent these limitations, we make use of DropArray, a recent technology that allows high retention of weakly adherent cells and suspension cells. DropArray offers the competitive advantage of maintaining the classic high throughput format of microtiter plates while reducing classic microwell volume by up to 90% by using a drop format. Here, we present a mitotic index cell-based assay using the mitosis marker phospho histone H3 at serine 10 on a DropArray 384-well plate format. Dose-response curve analysis of the mitotic index assay with an antimitotic drug (docetaxel) on DropArray is presented that shows an effective dosage compared to previous established results similar to those obtained with conventional microtiter plates. The mitotic index assay with DropArray showed a Z-factor >0.6. Our results validate DropArray as a suitable platform for high throughput screening for compounds affecting mitosis or the cell cycle.