Immunostaining of intact C. elegans using polyacrylamide embedding

STAR Protoc. 2023 Mar 17;4(1):101956. doi: 10.1016/j.xpro.2022.101956. Epub 2023 Feb 9.

Abstract

A major barrier to immunostaining Caenorhabditis elegans is the permeabilization of the worm's cuticle without distorting or damaging its body. We present here a gel-based immobilization protocol for fixed worms coupled with chemical and enzymatic permeabilization. The permeabilization is followed by antibody staining and fluorescent imaging. This protocol can be modified for different fixatives, permeabilizing reagents, or molecular readouts. Unlike previous immunostaining approaches, such as freeze cracking or dissection, this protocol enables immunostaining across the whole body of a well-preserved C. elegans.

Keywords: Antibody; Cell Biology; Developmental biology; Microscopy; Model Organisms; Molecular Biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylic Resins*
  • Animals
  • Antibodies
  • Caenorhabditis elegans* / chemistry
  • Fixatives

Substances

  • polyacrylamide
  • Acrylic Resins
  • Fixatives
  • Antibodies