Whole-Mount Immunolocalization Procedure for Plant Female Meiocytes

Methods Mol Biol. 2020:2061:13-24. doi: 10.1007/978-1-4939-9818-0_2.

Abstract

Here we present an optimized protocol for immunolocalization of meiotic proteins during female meiosis in whole mount tissues. It ensures ovule morphology integrity and homogeneous reagent penetration. The method relies on paraformaldehyde tissue fixation, polyacrylamide embedding, tissue permeabilization, antibody incubation, counterstaining, and confocal microscopy analysis. This protocol has been used in diverse Arabidopsis ecotypes and in the legume Vigna unguiculata.

Keywords: Confocal microscopy; Female meiosis; Immunolocalization; Plant; Whole-mount.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / cytology
  • Arabidopsis / metabolism
  • Immunohistochemistry* / methods
  • Meiosis*
  • Microscopy, Confocal
  • Plant Cells / physiology*