Imaging tools for analysis of the ureteric tree in the developing mouse kidney

Luise Anne Cullen-McEwen, Richard James Young, Gabriel Fricout, Dominique Jeulin, Ian Steward Harper, Frank Costantini, John Frederick Bertram

Research output: Contribution to journalArticleResearchpeer-review

2 Citations (Scopus)

Abstract

The structure of the ureteric tree in developing mouse and rat kidneys has previously been quantified in two dimensions. While this type of analysis may provide evidence of changes in ureteric growth, these measurements are effectively inaccurate, as the ureteric tree is a three-dimensional (3D) object. Here we describe a method for measuring the ureteric tree in three dimensions. This technique involves (1) culture of the metanephric kidney at embryonic day 12 (mouse) or 14 (rat), (2) whole-mount immunofluorescence to selectively stain ureteric tree epithelium, (3) confocal microscopy to obtain a complete Z series through the ureteric tree, and (4) image analysis algorithms to binarize, skeletonize, and measure individual branch lengths in 3D. This method has been extended to analysis of the same ureteric tree over time (4D). The results obtained provide accurate and precise quantitation of ureteric tree growth in the developing mouse or rat kidney.
Original languageEnglish
Pages (from-to)305-320
Number of pages16
JournalMethods in Molecular Biology
Volume1075
DOIs
Publication statusPublished - 2014

Keywords

  • Confocal
  • Development
  • Image processing
  • Kidney
  • Measurement
  • Metanephros
  • Three-dimensional
  • Ureteric tree

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