MouseHouseApp Blog

In-situ Hybridization

Posted by Henry Kimball

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After working out of MassChallenge in Boston — housing 128 startups all in one awesome space, I’m finally back in Chicago and back to work at Moskowitz Lab! 

As November starts to roll in with the cold weather, Moskowitz lab is kicking it into full-gear!  Right now we’re in the middle of a whole mount in situ hybridization. While we do have many, many washes in store for us, this lengthy process with hopefully be worth the wait (and hard work)! 

Here I’ve outlined what process we are doing — In situ hybridization (ISH). It’s used to to locate specific segments of nucleic acid (DNA or RNA sequence) in a part of a tissue. These nucleic acids can be detected by applying a complementary strand of nucleic acid and then attaching reporter molecule to the complementary strand.

Whole mount ISH means the tissue is small enough that the whole tissue can be used such as what we are using — mice embryos. Remember not to confuse it with immunohistochemisty, which is used for locating proteins in tissues. Our case of using RNA probes, or Riboprobes, for our ISH measures and localizes these RNAs (mRNAs, lncRNAs and miRNAs) within this whole mount tissue. Also, riboprobes allow us to determine the degree of gene expression.

The main applications of ISH include discovering morphology and population structure of microorganisms in microbiology, profiling pathogens and abnormal gene expression in pathology, profiling gene expression in embryonic tissues in developmental biology, counting the number and noting visual appearance of the chromosomes in the cell nuclei of an organism or species in karyotyping and phylogenetic analysis, and mapping clones on chromosomes as well as direct assignment through physical mapping.

Read more about it here:

http://www.ncbi.nlm.nih.gov/genome/probe/doc/TechISH.shtml

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