Double labeling for whole-mount in situ hybridization in mouse.

Combined fluorescence in situ hybridization and immunocytochemistry double labeling in whole-mount Manduca sexta tissue.

Whole-mount in situ hybridization to mouse embryos.

The mouse is well-established as the major animal model for the study of mammalian development. Rapid progress in large-scale cDNA and also genomic sequencing projects is identifying new mouse genes at an unprecedented rate. As a first step toward understanding the function of these novel genes, it is important to determine their developmental expression pattern. Here we provide a reliable, sen...

Whole mount in situ hybridization of E8.5 to E11.5 mouse embryos.

Whole mount in situ hybridization is a very informative approach for defining gene expression patterns in embryos. The in situ hybridization procedures are lengthy and technically demanding with multiple important steps that collectively contribute to the quality of the final result. This protocol describes in detail several key quality control steps for optimizing probe labeling and performanc...

Whole-mount in situ hybridization of RNA probes to plant tissues.

INTRODUCTIONThe labeling of entire plantlets (whole-mount labeling) is very easy, but it can cause substantial artifacts. Because whole plants are labeled, certain tissues are not readily accessible to the probes, whereas others can often stain nonspecifically. In the hands of the author, for instance, the elongation zone of the root is labeled even in negative controls, whereas mature parts of...

دورگه‌سازی در محل؛ اصول و کاربردها : مقاله مروری

In situ hybridization (ISH) is a method that uses labeled complementary single strand DNA or RNA to localize specific DNA or RNA sequences in an intact cell or in a fixed tissue section. The main steps of ISH consist of: probe selection, tissue or sample preparation, pre-hybridization treatment, hybridization and washing, detection and control procedure. Probe selection is one of the important ...

High-Resolution Whole-Mount In Situ Hybridization Using Quantum Dot Nanocrystals

The photostability and narrow emission spectra of nanometer-scale semiconductor crystallites (QDs) make them desirable candidates for whole-mount fluorescent in situ hybridization to detect mRNA transcripts in morphologically preserved intact embryos. We describe a method for direct QD labeling of modified oligonucleotide probes through streptavidin-biotin and antibody-mediated interactions (an...