Immunofluorescence Microscopy

Diane C. Shakes, David M. Miller, Michael L. Nonet

Research output: Chapter in Book/Report/Conference proceedingChapter

13 Scopus citations

Abstract

Immunofluorescence microscopy is a powerful technique that is widely used by researchers to assess both the localization and endogenous expression levels of their favorite proteins. The application of this approach to C. elegans, however, requires special methods to overcome the diffusion barrier of a dense, collagen-based outer cuticle. This chapter outlines several alternative fixation and permeabilization strategies for overcoming this problem and for producing robust immunohistochemical staining of both whole animals and freeze-fractured samples. In addition, we provide an accounting of widely used antibody reagents available to the research community.We also describe several approaches aimed at reducing non-specific background often associated with immunohistochemical studies.Finally, we discuss a variety of approaches to raise antisera directed against C. elegans antigens.

Original languageEnglish
Title of host publicationMethods in Cell Biology
PublisherAcademic Press Inc.
Pages35-66
Number of pages32
DOIs
StatePublished - 2012

Publication series

NameMethods in Cell Biology
Volume107
ISSN (Print)0091-679X

Keywords

  • Immunofluorescence
  • Permeabilization
  • Polyclonal
  • Polyclonal antibodies
  • Posttranslational
  • Transgenic

Fingerprint Dive into the research topics of 'Immunofluorescence Microscopy'. Together they form a unique fingerprint.

  • Cite this

    Shakes, D. C., Miller, D. M., & Nonet, M. L. (2012). Immunofluorescence Microscopy. In Methods in Cell Biology (pp. 35-66). (Methods in Cell Biology; Vol. 107). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-394620-1.00002-3