DGIST: A standardized image-based spatial quantification workflow for neurogenesis in the dentate gyrus.

The adult hippocampus generates new neurons in the dentate gyrus (DG), where neural stem cells (NSCs) in the subgranular zone differentiate into neuronal and astroglial lineages. Accurate quantification of adult NSCs is critical for assessing neurogenic potential, yet most current methods rely on manual, density-based measurements in limited regions of the DG. These approaches may introduce bias due to morphological variability among tissue sections. To overcome these challenges, we developed the Dentate Gyrus Image-based Spatial quantification Tool (DGIST), an ImageJ/Fiji-based workflow that integrates BioVoxxel and MorphoLibJ plug-ins for cell-based quantification throughout the entire DG. Applied to in vivo datasets, DGIST quantifies age-dependent changes in marker-defined NSC and astroglial populations. Our results demonstrate that DGIST provides a reproducible method for quantifying marker-defined populations in the adult neurogenic niche.