Multi-modal label-free imaging of cellular metabolism and oxidative stress in 3D brain tissue models.

ABSTRACT: Label-free, two-photon fluorescence imaging enables non-destructive, single-cell metabolic assessments that capture metabolic reprogramming and heterogeneity. Here, we use intensity and lifetime imaging to report on cell metabolic function and dynamic interactions in 3D engineered brain tissue models, comprising human induced neural stem cells, astrocytes and microglia. We rely on spectral imaging to define excitation/emission profiles of key fluorophores, including NAD(P)H, lipoamide dehydrogenase, FAD, and lipofuscin. These profiles inform a multi-wavelength imaging strategy that accounts for lipofuscin and spectral overlap, ensuring accurate redox state, mitochondrial fragmentation, and NAD(P)H bound state analysis. Intensity imaging over 11 weeks reveals metabolic differences between n

SUBMITTER: Zhang Y 

PROVIDER: S-EPMC12673076 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature