Biology subjects

Cicerone, M.

Publications and source records attributed to Cicerone, M..

3 recordsLinked to original sources

bioRxiv2026

Multiparametric in vivo mapping reveals tissue-specific mitochondrial aging trajectories

Mitochondrial dysfunction is a hallmark of aging, yet how mitochondrial states are remodeled across tissues and subcellular compartments in vivo remains elusive. Progress has been limited, in part, because mitochondrial physiology is highly sensitive to experimental perturbations, underscoring the need for minimally disruptive measurement strategies. Here, we establish a tissue-resolved, in vivo framework for the quantitative analysis of mitochondrial states in live, intact Caenorhabditis elegans without confounding effects from mounting-induced hypoxia. This platform couples two-photon fluorescence lifetime imaging microscopy (2p-FLIM) with a custom segmentation pipeline, MitoSLIT, to track functional and structural features across multiple tissues and single neurons. By integrating membrane potential-associated TMRM intensity, lifetime-based microenvironmental metrics, and morphological descriptors, we uncover localized metabolic heterogeneity masked by conventional intensity analysis. Leveraging this framework, we mapped physiological aging against mitochondrial shifts induced by acute stress and fission-fusion mutations. Our analyses reveal that mitochondrial aging is highly tissue-specific, executing distinct trajectories across cell types. Extending the framework to genetically identified neurons revealed age-dependent divergence between somatic and axonal mitochondrial states, accompanied by structural remodeling and a late shift in optical redox ratio. Together, our findings demonstrate that mitochondrial populations do not converge on a uniform bioenergetic endpoint during aging, but rather follow highly compartmentalized, tissue-specific spatiotemporal trajectories in vivo.

physiology↗

bioRxiv2026

Deep-ultraviolet microscopy reveals biomolecular spatiotemporal intracellular dynamics

Intracellular dynamics span a broad range of time scales and biomolecular processes, offering insights into cell health, functional state, phenotype, and response to external perturbations. Several label-free optical imaging approaches have been used to capture intracellular dynamics but are limited by spatiotemporal resolution and biomolecular specificity required to distinguish unique subcellular and metabolic processes. In this work, we demonstrate deep-ultraviolet (UV) microscopy as a powerful, label-free, high-resolution approach for quantifying multiscale intracellular dynamics with biomolecular specificity. By leveraging power spectral analysis and phasor analysis, we capture multiscale intracellular dynamics and analyze their UV wavelength-dependent behavior predicated by the absorption of different endogenous biomolecules. We apply this technique to prostate epithelial cell lines of increasing malignancy and reveal quantitative differences in dynamic intracellular activity that correlate with increased metabolic and organelle activity between phenotypes. Furthermore, we elucidate the molecular identities of structures and activity measured via UV dynamics with broad-band coherent anti-Stokes Raman scattering spectroscopy and fluorescence microscopy. We identify lipid-specific structures, and mitochondrial-specific dynamics, among other biomolecular-specific dynamic behaviors. Together, this study demonstrates deep-UV microscopy as a powerful imaging platform for probing spatial and temporally variant intracellular dynamics with biomolecular specificity, with broad implications for cell phenotyping, tissue pathology, and studying new dynamic subcellular processes.

cell biology↗

bioRxiv2026

Hydrogel Fiber Endomicroscopy

Multimode fibers enable minimally invasive, high-resolution imaging through ultrathin probes, thereby enhancing diagnostic precision and facilitating real-time monitoring in delicate anatomical regions. In this work, we introduce HYFEN, a hydrogel-based endomicroscopic imaging platform for flexible, biocompatible, and subcellular-scale fluorescence microscopy. HYFEN leverages the unique properties of hydrogel materials, adaptive optics, and pixel-wise image enhancement to address challenges associated with silica-based fibers, including mode scrambling, limited field of view, and mechanical rigidity. The technique achieves precise mode threading, rapid diffraction-limited focusing at kilohertz speeds, and high-fidelity fluorescence signal acquisition with subcellular resolution. Notably, the approach extends fluorescence imaging under enhanced fiber dimensions and bending conditions that are unachievable with conventional modalities. Together, these advances establish HYFEN as a versatile platform for next-generation biointerfacing and minimally invasive imaging across biomedical and clinical settings.

bioengineering↗