Embedding Adult Drosophila Heads in Paraffin: Preparing Tissue for Histology

In This Article

Abstract

Source: Sunderhaus, E. R., Kretzschmar, D. Mass Histology to Quantify Neurodegeneration in Drosophila .J. Vis. Exp. (2016).

This video describes how to embed Drosophila heads in paraffin using the collar method, a technique that prepares fly heads for serial sectioning. Paraffin sections are used for histological examinations of the brain's anatomy and for the assessment of neurodegeneration. The featured protocol demonstrates how multiple flies can be processed in one preparation, thereby increasing time-efficiency and minimizing experimental artifacts.

Protocol

This protocol is an excerpt from Sunderhaus and Kretzschmar, Mass Histology to Quantify Neurodegeneration in Drosophila, J. Vis. Exp. (2016).

1. Fixing the Head on Collars and Embedding in Paraffin

NOTE: All of the steps in the fixation process should be done in a fume hood. Methylbenzoate, while not posing a health risk, has a highly distinct odor, which can be overwhelming if not handled in a fume hood.

Before anesthetizing the flies, make up 50 mL of Carnoy solution by adding 15 mL of chloroform and 5 mL of glacia....

Access restricted. Please log in or start a trial to view this content.

Results

Mechanical deformation study; microstructure schematic; fluorescence imaging of cellular alignment.
Figure 1: Paraffin Serial Sections. A) Using the collar method, experimental and control flies can be processed as one sample by threading them onto one collar. Eyeless sine oculis flies are inserted for orientation (arrows). B) Sch.......