Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy

Carrie Phillips, L. J. Arend, A. J. Filson, D. J. Kojetin, J. L. Clendenon, S. Fang, Kenneth Dunn

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Developing mammalian embryonic kidney becomes progressively more elaborate as the ureteric bud branches into undifferentiated mesenchyme. Morphological perturbations of nephrogenesis, such as those seen in inherited renal diseases or induced in transgenic animals, require careful and often tedious documentation by multiple methodologies. We have applied a relatively quick and simple approach combining two-photon microscopy and advanced three-dimensional (3-D) imaging techniques to visualize and evaluate these complex events. As compared with laser confocal microscopy, two-photon microscopy offers superior optical sectioning deep into biological tissues, permitting analysis of large, heterogeneous, 3-D structures such as developing mouse kidney. Embryonic and newborn mouse kidneys were fluorescently labeled with lectins, phalloidin, or antibody. Three-dimensional image volumes were then collected. The resulting volume data sets were processed using a novel 3-D visualization technique. Reconstructed image volumes demonstrate the dichotomous branching of ureteric bud as it progresses from a simple, symmetrical structure into an elaborate, asymmetrical collecting system of multiple branches. Detailed morphology of in situ cysts was elucidated in a transgene-induced mouse model of polycystic kidney disease. We expect this integration of two-photon microscopy with advanced 3-D image analysis will provide a powerful tool for illuminating a variety of complex developmental processes in multiple dimensions.

Original languageEnglish
Pages (from-to)49-55
Number of pages7
JournalAmerican Journal of Pathology
Volume158
Issue number1
StatePublished - 2001

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Three-Dimensional Imaging
Photons
Microscopy
Kidney
Confocal Microscopy
Phalloidine
Polycystic Kidney Diseases
Genetically Modified Animals
Mesoderm
Transgenes
Lectins
Documentation
Cysts
Antibodies

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Phillips, C., Arend, L. J., Filson, A. J., Kojetin, D. J., Clendenon, J. L., Fang, S., & Dunn, K. (2001). Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy. American Journal of Pathology, 158(1), 49-55.

Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy. / Phillips, Carrie; Arend, L. J.; Filson, A. J.; Kojetin, D. J.; Clendenon, J. L.; Fang, S.; Dunn, Kenneth.

In: American Journal of Pathology, Vol. 158, No. 1, 2001, p. 49-55.

Research output: Contribution to journalArticle

Phillips, C, Arend, LJ, Filson, AJ, Kojetin, DJ, Clendenon, JL, Fang, S & Dunn, K 2001, 'Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy', American Journal of Pathology, vol. 158, no. 1, pp. 49-55.
Phillips C, Arend LJ, Filson AJ, Kojetin DJ, Clendenon JL, Fang S et al. Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy. American Journal of Pathology. 2001;158(1):49-55.
Phillips, Carrie ; Arend, L. J. ; Filson, A. J. ; Kojetin, D. J. ; Clendenon, J. L. ; Fang, S. ; Dunn, Kenneth. / Three-dimensional imaging of embryonic mouse kidney by two-photon microscopy. In: American Journal of Pathology. 2001 ; Vol. 158, No. 1. pp. 49-55.
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