Mounting Adult Drosophila Brains: A Method to Prepare Slides for Confocal Imaging

In This Article

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 a method of mounting intact brains from adult flies under a bridge slide and shows the protocol performed on immunostained brains for fluorescent microscopy.

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. Mounting Adult D. melanogaster Brains onto Microscope Slides and Imaging

Build a "bridge" slide. Position two "base" coverslips roughly 1 cm apart on a positively charged slide. Ensure that the positively charged side of the slide is facing up. Adhere the coverslips to the slide with fingernail polish as shown in Figure 1A. It is usually ....

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

Results

Microscopy cover slip setup diagram, Vectashield filling for sample preservation.
Figure 1: Mounting brains in preparation for immunofluorescence imaging. (A) Brains are mounted on SuperFrost Plus slides with a bridge cover to prevent flattening of the brains. Two "base" cover slips are adhered to a Superfrost Plus positively-charged slide with clear finge.......