Mounting Adult Drosophila Brains: A Method to Prepare Slides for Confocal Imaging
Published: April 30, 2023
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 sl…
Representative Results
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 fingernail polish and dissected brains are then placed on the slide between them. A clear "bridge" cover slip is pla…
Materials
| Microdissection forceps/tweezers | Ted Pella | 505-NM | |
| VectaShield | Vector Labs | H-1000 | |
| SuperFrost Plus Slides | ThermoFisher | 99-910-01 | |
| Coverslips | ThermoFisher | 12-553-454 | |
| fingernail polish | Electron Microscope Services (EMS) | 72180 | |
| stereomicroscope | Leica S6D with KL300 LED light source | ||
| Kimwipe | Fisher | 06-666 |
Cite This Article
Mounting Adult Drosophila Brains: A Method to Prepare Slides for Confocal Imaging. J. Vis. Exp. (Pending Publication), e20106, doi: (2023).