Whole Mount In Situ
Whole Mount In Situ - Related Videos
08:00
Whole-Mount In Situ Hybridization
0 Views •
2023
Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose...
12:31
Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization
0 Views •
Cited by 133 •
2009
Whole mount in situ hybridization is one of the most widely used techniques in developmental biology. Here, we present a high-resolution double fluorescent in situ hybridization protocol for analyzing the precise expression pattern of a single gene and for determining the overlap of the expression domains of two genes. We include a propidium iodide nuclear counter-stain to highlight tissue organization.
15:42
Double Whole Mount in situ Hybridization of Early Chick Embryos
0 Views •
Cited by 4 •
2008
This video demonstrates 2-color whole mount in situ hybridization, a method by which the spatial and temporal expression pattern of 2 different genes can be visualized in young chick embryos. This method was originally introduced by David Wilkinson, Domingos Henrique, Phil Ingham and David Ish -Horowicz.
07:33
A Whole Mount In Situ Hybridization Method for the Gastropod Mollusc Lymnaea stagnalis
0 Views •
Cited by 17 •
2016
The goal of this protocol is to provide users with a set of methods for the high-throughput decapsulation of Lymnaea stagnalis embryos and larvae in preparation for whole mount in situ hybridization, and for subsequent pre- and post-hybridization treatments.
10:06
High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
0 Views •
Cited by 24 •
2013
The zebrafish, a small tropical fish, has become a popular model for studying gene function during vertebrate development and disease. The temporal and spatial expression of target genes can be determined by in situ hybridization. Our improved protocol allows for the detection of low abundant transcripts with low non-specific background signal.
FAQs
A labeled complementary probe binds the matching messenger RNA sequence within the preserved sample. Subsequent washing removes probe that has not hybridized, reducing nonspecific background and leaving signal where the target transcript is present. The resulting pattern reflects spatial distribution rather than simply total transcript abundance, allowing expression to be compared with visible anatomical structures.
Fixation preserves the embryo, tissue, or organism so its anatomical relationships remain available for interpretation, while permeabilization allows the probe to enter the sample. These steps must support both structural preservation and probe access. Inadequate access can reduce detectable signal, whereas limited penetration becomes a particular concern when targets lie deep inside an intact specimen.
Whole-mount analysis preserves the specimen as an intact three-dimensional arrangement, making it possible to view expression domains across an entire embryo, tissue, or small organism. Section-based assays can provide higher local resolution in individual slices, but they may fragment spatial relationships. The choice therefore depends on whether global anatomical pattern or fine section-level detail is more important.
Both approaches convert probe binding into a visible spatial signal, but they present expression patterns differently. Colorimetric detection reveals labeled regions through a visible reaction, whereas fluorescent detection displays signal through fluorescence. In either case, interpretation depends on where signal appears relative to anatomy, developmental stage, and the method’s limits in penetration and resolution.
A typical workflow fixes the intact specimen, permeabilizes it, exposes it to a labeled complementary probe, washes away unbound probe, and then performs colorimetric or fluorescent detection. Examining the resulting signal across the preserved anatomy connects transcript location with structure. Careful interpretation is especially important when probe access or signal visibility varies within the specimen.
The method is useful when researchers need to relate gene expression to embryonic patterning, cell fate, or organ formation across an intact specimen. Because the spatial pattern remains associated with developing anatomy, investigators can examine where a transcript is detected at particular developmental stages. The same principle also supports analysis of tissue organization and disease models.