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Live-cell Imaging of Endocytic Transport using Functionalized Nanobodies in Cultured Cells
Published on: October 17, 2025
Quantifying endocytosis in vivo using intravital two-photon microscopy
Ruben M Sandoval1, Bruce A Molitoris
1Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA.
Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2008
Summary
Multiphoton microscopy allows researchers to track molecular uptake and metabolism in kidney cells with subcellular detail. This intravital technique offers new insights into kidney physiology and disease in preclinical models.
Area of Science:
- Cell Biology
- Physiology
- Biomedical Imaging
Background:
- Multiphoton microscopy has advanced, enabling quantitative, cell-specific studies of dynamic events like endocytosis within intact organs.
- This technology offers superior spatial, temporal resolution, and sensitivity for tracking molecules and their cellular fates.
Purpose of the Study:
- To utilize intravital multiphoton microscopy to study kidney proximal tubule cell uptake, distribution, and metabolism.
- To quantify the cellular handling of fluorescently labeled molecules such as folic acid, gentamicin, and small interfering ribonucleic acid (siRNA).
Main Methods:
- Intravital multiphoton microscopy was employed in preclinical models.
- Simultaneous use of up to three fluorophores allowed for volumetric data quantification.
- Cell-specific tracking of molecular uptake, intracellular trafficking, and metabolism was performed.
Main Results:
- The study successfully quantified kidney proximal tubule cell uptake and intracellular distribution of labeled folic acid, gentamicin, and siRNA.
- The technique provided cell-specific insights into molecular processing within the complex kidney microenvironment.
Conclusions:
- Intravital multiphoton microscopy is a powerful tool for studying kidney cell physiology and disease pathophysiology in preclinical settings.
- Tissue penetration remains a limitation for direct clinical translation, but preclinical insights are significant.
