Quantitative intravital microscopy using a Generalized Polarity concept for kidney studies

Weiming Yu, Ruben M. Sandoval, Bruce Molitoris

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

In this article, we describe a ratiometric intravital two-photon microscopy technique for studying glomerular permeability and differences in proximal tubule cell reabsorption. This quantitative approach is based on the Generalized Polarity (GP) concept, in which the intensity difference between two fluorescent molecules is normalized to the total intensity produced by the two dyes. After an initial intravenous injection of a mixture of 3-, 40-, and 70-kDa fluorescently labeled dextrans into live Munich-Wistar-Frömter (MWF) rats, we were able to monitor changes in the GP values between any two dyes within local regions of the kidney, including the glomerulus, Bowman's capsule, proximal tubule lumens and proximal tubule cells, and individual capillary vessels. We were able to quantify accumulations of different dextrans in the Bowman's space and in tubular lumens as well as reabsorption by proximal tubular cells at different time points in the same rat. We found that for 6- to 8-wk-old MWF rats that developed spontaneous albuminuria, the 40- and 70-kDa dextrans, with hydrodynamic radii larger than albumin, were differentially filtered, but both were able to pass the glomerular filtration barrier and enter into the urinary space of the Bowman's capsule within a few seconds after intravenous infusion. Using GP image analysis, we found that negatively charged dextrans of both 40 and 70 kDa were better reabsorbed by the proximal tubule cells than the neutrally charged 40-kDa dextran. These results demonstrate the potential power of the GP imaging technique for quantitative studies of glomerular filtration and tubular reabsorption.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume289
Issue number5 58-5
DOIs
StatePublished - Nov 2005

Fingerprint

Dextrans
Kidney
Bowman Capsule
Rats
Capsules
Wistar Rats
Coloring Agents
Glomerular Filtration Barrier
Kidney Glomerulus
Albuminuria
Hydrodynamics
Photons
Intravenous Infusions
Intravenous Injections
Image analysis
Intravital Microscopy
Microscopy
Albumins
Permeability
Microscopic examination

Keywords

  • Charge selectivity
  • Glomerular permeability
  • Multiphoton
  • Tubular reabsorption
  • Two-photon excitation

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Quantitative intravital microscopy using a Generalized Polarity concept for kidney studies. / Yu, Weiming; Sandoval, Ruben M.; Molitoris, Bruce.

In: American Journal of Physiology - Cell Physiology, Vol. 289, No. 5 58-5, 11.2005.

Research output: Contribution to journalArticle

@article{8ca910635d5f4ecbbdee5b4a70da96fc,
title = "Quantitative intravital microscopy using a Generalized Polarity concept for kidney studies",
abstract = "In this article, we describe a ratiometric intravital two-photon microscopy technique for studying glomerular permeability and differences in proximal tubule cell reabsorption. This quantitative approach is based on the Generalized Polarity (GP) concept, in which the intensity difference between two fluorescent molecules is normalized to the total intensity produced by the two dyes. After an initial intravenous injection of a mixture of 3-, 40-, and 70-kDa fluorescently labeled dextrans into live Munich-Wistar-Fr{\"o}mter (MWF) rats, we were able to monitor changes in the GP values between any two dyes within local regions of the kidney, including the glomerulus, Bowman's capsule, proximal tubule lumens and proximal tubule cells, and individual capillary vessels. We were able to quantify accumulations of different dextrans in the Bowman's space and in tubular lumens as well as reabsorption by proximal tubular cells at different time points in the same rat. We found that for 6- to 8-wk-old MWF rats that developed spontaneous albuminuria, the 40- and 70-kDa dextrans, with hydrodynamic radii larger than albumin, were differentially filtered, but both were able to pass the glomerular filtration barrier and enter into the urinary space of the Bowman's capsule within a few seconds after intravenous infusion. Using GP image analysis, we found that negatively charged dextrans of both 40 and 70 kDa were better reabsorbed by the proximal tubule cells than the neutrally charged 40-kDa dextran. These results demonstrate the potential power of the GP imaging technique for quantitative studies of glomerular filtration and tubular reabsorption.",
keywords = "Charge selectivity, Glomerular permeability, Multiphoton, Tubular reabsorption, Two-photon excitation",
author = "Weiming Yu and Sandoval, {Ruben M.} and Bruce Molitoris",
year = "2005",
month = "11",
doi = "10.1152/ajpcell.00197.2005",
language = "English",
volume = "289",
journal = "American Journal of Physiology",
issn = "0193-1857",
publisher = "American Physiological Society",
number = "5 58-5",

}

TY - JOUR

T1 - Quantitative intravital microscopy using a Generalized Polarity concept for kidney studies

AU - Yu, Weiming

AU - Sandoval, Ruben M.

AU - Molitoris, Bruce

PY - 2005/11

Y1 - 2005/11

N2 - In this article, we describe a ratiometric intravital two-photon microscopy technique for studying glomerular permeability and differences in proximal tubule cell reabsorption. This quantitative approach is based on the Generalized Polarity (GP) concept, in which the intensity difference between two fluorescent molecules is normalized to the total intensity produced by the two dyes. After an initial intravenous injection of a mixture of 3-, 40-, and 70-kDa fluorescently labeled dextrans into live Munich-Wistar-Frömter (MWF) rats, we were able to monitor changes in the GP values between any two dyes within local regions of the kidney, including the glomerulus, Bowman's capsule, proximal tubule lumens and proximal tubule cells, and individual capillary vessels. We were able to quantify accumulations of different dextrans in the Bowman's space and in tubular lumens as well as reabsorption by proximal tubular cells at different time points in the same rat. We found that for 6- to 8-wk-old MWF rats that developed spontaneous albuminuria, the 40- and 70-kDa dextrans, with hydrodynamic radii larger than albumin, were differentially filtered, but both were able to pass the glomerular filtration barrier and enter into the urinary space of the Bowman's capsule within a few seconds after intravenous infusion. Using GP image analysis, we found that negatively charged dextrans of both 40 and 70 kDa were better reabsorbed by the proximal tubule cells than the neutrally charged 40-kDa dextran. These results demonstrate the potential power of the GP imaging technique for quantitative studies of glomerular filtration and tubular reabsorption.

AB - In this article, we describe a ratiometric intravital two-photon microscopy technique for studying glomerular permeability and differences in proximal tubule cell reabsorption. This quantitative approach is based on the Generalized Polarity (GP) concept, in which the intensity difference between two fluorescent molecules is normalized to the total intensity produced by the two dyes. After an initial intravenous injection of a mixture of 3-, 40-, and 70-kDa fluorescently labeled dextrans into live Munich-Wistar-Frömter (MWF) rats, we were able to monitor changes in the GP values between any two dyes within local regions of the kidney, including the glomerulus, Bowman's capsule, proximal tubule lumens and proximal tubule cells, and individual capillary vessels. We were able to quantify accumulations of different dextrans in the Bowman's space and in tubular lumens as well as reabsorption by proximal tubular cells at different time points in the same rat. We found that for 6- to 8-wk-old MWF rats that developed spontaneous albuminuria, the 40- and 70-kDa dextrans, with hydrodynamic radii larger than albumin, were differentially filtered, but both were able to pass the glomerular filtration barrier and enter into the urinary space of the Bowman's capsule within a few seconds after intravenous infusion. Using GP image analysis, we found that negatively charged dextrans of both 40 and 70 kDa were better reabsorbed by the proximal tubule cells than the neutrally charged 40-kDa dextran. These results demonstrate the potential power of the GP imaging technique for quantitative studies of glomerular filtration and tubular reabsorption.

KW - Charge selectivity

KW - Glomerular permeability

KW - Multiphoton

KW - Tubular reabsorption

KW - Two-photon excitation

UR - http://www.scopus.com/inward/record.url?scp=26844579948&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=26844579948&partnerID=8YFLogxK

U2 - 10.1152/ajpcell.00197.2005

DO - 10.1152/ajpcell.00197.2005

M3 - Article

C2 - 16033906

AN - SCOPUS:26844579948

VL - 289

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0193-1857

IS - 5 58-5

ER -