Poster Presentation 30th Lorne Cancer Conference 2018

Tracking cell division and viability during in vitro drug treatment using fluorescent time-lapse imaging. (#261)

Kristy L Shield-Artin , Gwo Yaw Ho , Elizabeth Kyran , Holly Barker , Matthew Wakefield , Clare Scott

Testing drug treatments in vitro is a key aspect of many preclinical models. Predominantly, single or serial end-point viability assays are used to determine efficacy of a drug. However, traditional endpoint viability assays cannot easily determine time dependent effects of treatment on cell division and cell death. Thus, alternative time-lapse monitoring options were investigated. Using what was available, an incubated Zeiss Axio-Observer widefield microscope with automated stage and the PerkinElmer Opera Phenix, we utilized a new live-cell compatible far-red DNA dye and captured regular images over 48h-72h. Finding a good non-toxic DNA dye that didn’t require UV-excitation (DNA damaging) was a key requirement for these studies. Initially, images taken at 90min intervals with the Axio-Observer and 20x Objective were used to confirmed that untreated cells could be tracked through multiple cell divisions with no observable defects. Furthermore, nuclear morphology was sufficient to identify mitotic cells prior to division. To improve throughput, plates were set up to be imaged on the Opera Phenix every 6h. Nuclei were counted across multiple fields per well and changes over time assessed. Importantly, when dyed cells and untreated cells were directly compared using an endpoint viability assay, no adverse effect was observed  when using the dye . Furthermore, the relative cell counts derived from the images were closely correlated to results from endpoint viability assays on the same cultures. This work illustrates that fluorescent labeling and time-lapse imaging of cells in vitro is possible without the need to transfect/transduce cells. A prospect that is enticing for sensitive cell lines and precious primary cultures.