Evaluating Tubulin Integrity and Cell Morphology

Imaging experiments were performed using an Operetta CLS high-content imaging device (PerkinElmer, Hamburg, Germany), and analysed with Harmony 4.6 software (PerkinElmer). To assess the tubulin integrity, we acquired HeLa cells images using 63× magnification, taking 25 fields per sample in biological and technical triplicates. Data were analysed using the following building blocks: 1—Find Nuclei, 2—Find Cytoplasm (Tubulin+). IC₅₀ was obtained by counting the number of nuclei in each field. Cell dimensions were assessed using the following building blocks: 1—Find Nuclei, 2—Find Cytoplasm (Tubulin+), 3—Calculate Morphology properties (Area), as previously described.^37–39 (link) IC₅₀s were calculated using the number of cells detected in every well as indexing parameter by Harmony 4.6 software as described here.⁴⁰ (link)