Quantifying endocytosis in vivo using intravital two-photon microscopy

Ruben M. Sandoval, Bruce Molitoris

Research output: Chapter in Book/Report/Conference proceedingChapter

20 Citations (Scopus)

Abstract

The recent introduction of multiphoton microscopy coupled with advances in optics, computer sciences, designer fluorophores, molecular labeling, and previously developed physiologic approaches have empowered investigators to quantitatively study the cell-specific dynamic events, such as endocytosis, within a functioning organ with subcellular resolution. This rapidly emerging field of investigation, with superior spatial and temporal resolution and high sensitivity, enables investigators to track molecules and determine their mode of cellular uptake, intracellular trafficking, and metabolism in a cell-specific fashion in complex heterogeneous organs such as the kidney with repeated determinations possible over a prolonged period of time. This approach is enhanced by the ability to obtain and quantify volumetric data with using up to three different fluorophores simultaneously. We have utilized this intravital approach to understand and quantify kidney proximal tubule cell uptake and intracellular distribution and metabolism of fluorescently labeled molecules, including folic acid, gentamicin, and small interfering ribonucleic acid (siRNA). Limitations of this technique include tissue penetration, which is the major barrier to successful clinical utilization of this technology. However, its use in preclinical animal models offers new insight into physiologic processes and the pathophysiology and treatment of disease processes.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages389-402
Number of pages14
Volume440
ISBN (Print)9781588298652
DOIs
StatePublished - 2008

Publication series

NameMethods in Molecular Biology
Volume440
ISSN (Print)10643745

Fingerprint

Endocytosis
Photons
Microscopy
Research Personnel
Proximal Kidney Tubule
Gentamicins
Folic Acid
Animal Models
RNA
Technology
Kidney

Keywords

  • Aminoglycosides
  • Dextrans
  • Gentamicin
  • Kidney imaging
  • Proximal tubule cells
  • SiRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Sandoval, R. M., & Molitoris, B. (2008). Quantifying endocytosis in vivo using intravital two-photon microscopy. In Methods in Molecular Biology (Vol. 440, pp. 389-402). (Methods in Molecular Biology; Vol. 440). Humana Press. https://doi.org/10.1007/978-1-59745-178-9_28

Quantifying endocytosis in vivo using intravital two-photon microscopy. / Sandoval, Ruben M.; Molitoris, Bruce.

Methods in Molecular Biology. Vol. 440 Humana Press, 2008. p. 389-402 (Methods in Molecular Biology; Vol. 440).

Research output: Chapter in Book/Report/Conference proceedingChapter

Sandoval, RM & Molitoris, B 2008, Quantifying endocytosis in vivo using intravital two-photon microscopy. in Methods in Molecular Biology. vol. 440, Methods in Molecular Biology, vol. 440, Humana Press, pp. 389-402. https://doi.org/10.1007/978-1-59745-178-9_28
Sandoval RM, Molitoris B. Quantifying endocytosis in vivo using intravital two-photon microscopy. In Methods in Molecular Biology. Vol. 440. Humana Press. 2008. p. 389-402. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-59745-178-9_28
Sandoval, Ruben M. ; Molitoris, Bruce. / Quantifying endocytosis in vivo using intravital two-photon microscopy. Methods in Molecular Biology. Vol. 440 Humana Press, 2008. pp. 389-402 (Methods in Molecular Biology).
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