Comparison of Trypan Blue and Fluorescence-Based Viability Detection Methods Via Morphological Observation
Determining cell viability is a vital component in many biological experiments that range from standard cell culture to the investigation of pharmacological agents on tumor cells. One
of the earliest and most common methods for measuring cell viability is the trypan blue (TB) exclusion assay [1, 2]. Over the last two decades, there have been various publications on
comparing TB exclusion and fluorescence-based cellular viability assays [1, 3-5]. Previous results have shown that in a time-course measurement, TB exclusion assays reported significantly higher viabilities when compared to fluorescence-based methods [4, 5]. These previous studies have only shown comparison results, but did not provide evidence on potential reasons for the differences. In this work, we performed a time-course study to compare cellular viability measured with TB and AO/PI using an automated image-based cytometry method (Cellometer, Nexcelom Bioscience).