Nuclear Morphology-Based Assessment of Cell Fates Induced by a Microtubule Targeting Agent as a Single Treatment or Combined with an Oncolytic Virus

Immunofluorescence microscopy allows for the quantitative assessment of cell fate at single-cell resolution. This is required to analyze heterogeneous cell populations, as the assessment of average values of given parameters does not faithfully describe distinct states of specific subpopulations. As a case in point, we describe a methodology for characterizing the effects of a microtubule targeting agent, 2-methoxestradiol (2ME2), on T24 human bladder cancer cells. We employ an immunofluorescence-based assessment of nuclear morphology, DNA content, and the intracellular distribution pattern of microtubules for the identification/classification of cells undergoing mitosis or mitotic slippage. When combined with imaging-based identification of cells expressing a nonstructural oncolytic virus protein, this approach enables the assessment of the potential for combined treatment with a microtubule targeting agent and an oncolytic virus (e.g., the Epizootic Hemorrhagic Disease Virus-Tel Aviv University, EHDV-TAU).