Asp Julia, Abramsson Alexandra, Betsholtz Christer
Department of Pathology, University of Goteborg, Goteborg, Sweden.
Methods Mol Biol. 2006;326:89-102. doi: 10.1385/1-59745-007-3:89.
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.
非放射性原位杂交为研究具有保留组织学信息的样本中的基因表达提供了独特的机会。这种方法不仅能够定位特定基因在组织中的表达位置，而且在许多情况下还能确定其在何种特定细胞类型中具有活性。在此，我们描述了目前用于小鼠胚胎冰冻切片或整体标本原位杂交的实验方案。所包含的方案描述了地高辛标记探针的合成、组织处理、探针与样本中表达的mRNA的杂交以及信号检测。
