An improved method to detect beta-galactosidase activity in transgenic mice: a post-staining procedure on paraffin embedded tissue sections

Transgenic Res. 1997 Jan;6(1):37-40. doi: 10.1023/a:1018401015497.

Abstract

The Escherichia coli beta-galactosidase gene is frequently used as a reporter gene in transgenic studies because its activity can be easily detected at the cellular level. Here we report a procedure for monitoring beta-galactosidase activity directly in tissue sections, which involves the use of a mixture of ethanol and poly-ethylene-glycol as a fixative (Kryofix) and a special paraffin characterized by a lower fusion point of 42 degrees C. After embedding and cutting, the sections are stained by the chromogenic substrate 5-bromo-4-chloro-3-indoyl-beta-D galactopyranoside (X-Gal). This procedure allows both the retention of a high level of beta-galactosidase activity and the preservation of good tissue morphology. Furthermore, it can be combined with immunohistochemical methods to detect other cellular components without compromising reporter gene detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arteries / anatomy & histology
  • Arteries / metabolism
  • Coloring Agents*
  • Embryo, Mammalian / anatomy & histology
  • Embryo, Mammalian / metabolism
  • Endothelium / anatomy & histology
  • Endothelium / metabolism
  • Escherichia coli / genetics
  • Genes, Reporter
  • Heart / anatomy & histology
  • Immunohistochemistry
  • Kidney / anatomy & histology
  • Kidney / metabolism
  • Lung / anatomy & histology
  • Lung / metabolism
  • Mice
  • Mice, Transgenic
  • Myocardium / metabolism
  • Paraffin Embedding / methods*
  • Rhombencephalon / anatomy & histology
  • Rhombencephalon / metabolism
  • Tissue Fixation / methods
  • Trigeminal Ganglion / anatomy & histology
  • Trigeminal Ganglion / metabolism
  • beta-Galactosidase / genetics*
  • beta-Galactosidase / metabolism*

Substances

  • Coloring Agents
  • beta-Galactosidase