Methods in Molecular Biology · Nov 13, 2026

An Integrated Pipeline for Quantifying Survival, Cell Morphology, and Nuclear Integrity During Chronological Aging of Fission Yeast

By Hairapatian K, Mallari K, Riffle T et al.

Chronological lifespan (CLS) assays in Schizosaccharomyces pombe are widely used to measure post-mitotic survival using colony-forming unit (CFU) readouts. Although CFU measurements provide a robust assessment of population-level viability, they do not directly capture cellular features that accompany survival decline during aging. Here, we describe an integrated experimental pipeline that enables the coordinated collection of three complementary readouts from a single CLS time course: CFU-based survival, automated quantification of cell morphology, and fluorescence-based assessment of nuclear integrity. During daily CLS sampling, aliquots from aging cultures are partitioned for viability plating, phase-contrast microscopy followed by automated image segmentation and statistical analysis, and ethanol fixation for DAPI staining. Automated morphology analysis yields cell-dimension distributions and normalized summary statistics aligned by aging day, while DAPI staining enables qualitative classification of nuclear morphology. This protocol allows population-level survival trajectories to be contextualized with single-cell morphological and nuclear-state measurements without requiring parallel experiments. The workflow is compatible with standard CLS assays and provides a reproducible framework for integrating viability, morphology, and nuclear integrity readouts during chronological aging in S. pombe.

doi.org/10.1007/978-1-0716-5703-4_5PubMed 43024201Biomarkers