In Situ Hybridization: Fruit Fly Embryos and Tissues

It is well known that transcript localization controls important biological processes, including cell fate determination, cell polarity, cell migration, morphogenesis, neuronal function, and embryonic axis specification. Thus, the sub-cellular visualization of transcripts in ‘their original place’ (in situ) is an important tool to infer and understand their trafficking, stability, translation, and biological functions. This has been made possible through the use of labeled ‘anti-sense’ probes that can be readily detected after hybridization to their ‘sense’ counterparts. The following is a series of protocols for conducting in situ hybridization in Drosophila embryos or tissues. These methods include standard alkaline phosphatase methods, as well as higher resolution and throughput variations using fluorescence-based probe detection. New modifications that enhance probe penetration and detection in various tissues are also provided. Curr. Protoc. Essential Lab. Tech. 4:9.3.1-9.3.24. © 2010 by John Wiley & Sons, Inc.