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Drug-Induced Reversible Lysosomal Changes Tracked in Live Cells by Holo-Tomographic Flow Cytometry
Daniele Pirone1, Michela Schiavo2,3,4, Giusy Giugliano1,3
1CNR-ISASI, Institute of Applied Sciences and Intelligent Systems "E. Caianiello", Via Campi Flegrei 34, Pozzuoli, Napoli 80078, Italy.
ACS Nano
|August 6, 2025
Summary
Holo-tomographic flow cytometry offers label-free 3D imaging of lysosomes in live cells. This breakthrough enables new biomarkers for lysosomal storage diseases and therapeutic strategy assessment.
Area of Science:
- Cell Biology
- Biophysics
- Medical Diagnostics
Background:
- Lysosomal storage diseases (LSDs) result from enzyme deficiencies, causing lysosomal dysfunction and cellular damage.
- Accurate assessment of lysosomal morphology and localization is crucial for understanding LSD pathology and developing treatments.
- Current methods have limitations in visualizing and quantifying lysosomal changes in real-time within live cells.
Purpose of the Study:
- To introduce and validate a novel holo-tomographic flow cytometry (HTFC) technique for label-free, high-content, 3D imaging of lysosomes.
- To establish quantitative biomarkers for lysosomal accumulation in LSD-affected cells using HTFC.
- To correlate 3D lysosomal architecture with the efficacy of therapeutic interventions for LSDs.
Main Methods:
- Utilized holo-tomographic flow cytometry (HTFC) for label-free, high-throughput 3D imaging of lysosomes in single live cells.
- Generated refractive index tomograms to measure and visualize cytoplasmic lysosomal aggregation.
- Performed experimental validation and computational analyses to link lysosomal architecture with therapeutic outcomes.
Main Results:
- Successfully identified and characterized lysosomes in live cells using HTFC, overcoming limitations of traditional methods.
- Developed quantitative biomarkers for lysosomal accumulation, enabling precise measurement of disease-related changes.
- Established a quantitative correlation between 3D lysosomal architecture and the effectiveness of genetic and pharmacological therapies.
Conclusions:
- HTFC provides a powerful, label-free tool for advancing lysosomal research and understanding LSDs.
- The developed quantitative biomarkers and 3D imaging capabilities can significantly improve LSD diagnostics.
- This technology holds promise for accelerating the development of targeted therapies and improving patient outcomes for lysosomal storage diseases.
