Drosophila Adult Brain Dissection: A Method in Fly Neurobiology

Abstract

Source: Kelly, S. M., et al. Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains. J. Vis. Exp. (2017).

This video describes how to dissect and isolate the adult Drosophila brain–a procedure necessary for visualizing the organ, which is otherwise obscured by the cuticle, eye, and tracheal tissue. The example protocol shows a detailed demonstration yielding high-quality preparations that can be used for immunostaining and imaging methods in Drosophila neurobiology.

Protocol

This protocol is an excerpt from Kelly et al., Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains, J. Vis. Exp. (2017).

1. Dissection Station Preparation

1. Position the stereomicroscope and light source with attached fiber optic goosenecks on a large benchtop. To promote steady hand movements and reduce hand "shake" while dissecting, it is essential that adequate hand and arm rest space is available around the microscope. Ensure that there is approximately 8 - 10 inches on either side of the mi....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Microdissection forceps/tweezers	Ted Pella	505-NM
Sylgard dishes	Living Systems Instrumentation	DD-50-S-BLK	Available from amazon.com
Na Phosphate Buffer monobasic	Sigma	S3139
Triton X 100	Sigma	X100-100ml
stereomicroscope	Leica S6D with KL300 LED light source
9-well dish (spot plate)	VWR	89090-482
35mm dish	Genesee Scientific	32-103
Sylgard	Fisher	50-366-794
Kimwipe	Fisher	06-666
PTN Buffer			0.1M NaPhosphate, pH 7.2, 0.1% Triton-X-100, Typically make up 0.5 L of 0.1M NaPhosphate buffer and aliquote 50ml at a time as needed