An improved method to detect beta-galactosidase activity in transgenic mice: a post-staining procedure on paraffin embedded tissue sections.
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.
Avé
P
P
Unité d'Histopathologie, Institut Pasteur, Paris, France.
Colucci-Guyon
E
E
Babinet
C
C
Huerre
M R
MR
D000818:Animals; D001158:Arteries / Q000033:anatomy & histology / Q000378:metabolism; D004396:Coloring Agents; D004622:Embryo, Mammalian / Q000033:anatomy & histology / Q000378:metabolism; D004727:Endothelium / Q000033:anatomy & histology / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D006321:Heart / Q000033:anatomy & histology; D007150:Immunohistochemistry; D007668:Kidney / Q000033:anatomy & histology / Q000378:metabolism; D008168:Lung / Q000033:anatomy & histology / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009206:Myocardium / Q000378:metabolism; D016612:Paraffin Embedding / Q000379:methods*; D012249:Rhombencephalon / Q000033:anatomy & histology / Q000378:metabolism; D016707:Tissue Fixation / Q000379:methods; D012668:Trigeminal Ganglion / Q000033:anatomy & histology / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism*
D004396:Coloring Agents; D001616:beta-Galactosidase