Unit

UNIT 9.3 In Situ Hybridization: Fruit Fly Embryos and Tissues

  1. Ronit Wilk1,2,
  2. Sreenivasa U.M. Murthy1,2,
  3. Haixu Yan1,2,
  4. Henry M. Krause1,2

Published Online: 1 DEC 2010

DOI: 10.1002/9780470089941.et0903s04

Current Protocols Essential Laboratory Techniques

Current Protocols Essential Laboratory Techniques

How to Cite

Wilk, R., Murthy, S. U., Yan, H. and Krause, H. M. 2010. In Situ Hybridization: Fruit Fly Embryos and Tissues. Current Protocols Essential Laboratory Techniques. 4:9.3:9.3.1–9.3.24.

Author Information

  1. 1

    Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada

  2. 2

    The Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada

Publication History

  1. Published Online: 1 DEC 2010
  2. Published Print: DEC 2010

Abstract

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.

Keywords:

  • in situ hybridization;
  • Drosophila;
  • embryos;
  • Drosophila;
  • larvae;
  • mRNA localization;
  • sub-cellular localization;
  • fluorescence;
  • tyramide;
  • FISH