Nonradioactive in situ hybridization on frozen sections and whole mounts.
Nonradioactive in situ hybridization offers a unique opportunity to study gene expression on samples with preserved histological information. This method makes it possible to locate not only where in a tissue a particular gene is expressed, but in many cases also in which specific cell type it is active. Here, we describe our current protocols for in situ hybridization on frozen sections or whole mounts of mouse embryos. The protocols included describe synthesis of a digoxigenin-labeled probe, tissue handling, hybridization of the probe to the mRNA expressed in the sample and signal detection.
Asp
Julia
J
Department of Pathology, University of Goteborg, Goteborg, Sweden.
Abramsson
Alexandra
A
Betsholtz
Christer
C
D000818:Animals; D004076:Digoxigenin / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D005629:Frozen Sections; D015870:Gene Expression; D006652:Histological Techniques; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D015335:Molecular Probes / Q000138:chemical synthesis; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011247:Pregnancy; D015347:RNA Probes / Q000138:chemical synthesis; D012333:RNA, Messenger / Q000378:metabolism
D015335:Molecular Probes; D015347:RNA Probes; D012333:RNA, Messenger; D004076:Digoxigenin