Protocol

Preparing Late Whole-Mount Drosophila Embryos for Immunostaining

This protocol was adapted from “Staining Tissues,” Chapter 6, in Using Antibodies by Ed Harlow and David Lane. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1999.

INTRODUCTION

The choice of fixation and permeabilization methods for examining flies depends on the stage of development. Early and late embryos are treated with a mixture of organic solvents, formaldehyde, and alcohols. The cuticles of late-stage embryos (17-22 hours or until hatching) are usually opened by sonication, as described here. Tissues from later stages of development are normally dissected by hand and then fixed and stained in a standard paraformaldehyde/detergent combination.

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