Drosophila Male Accessory Gland Dissection: A Method to Isolate Secondary Cells
Published: April 30, 2023
Abstract
Source: Immarigeon, C., et al. A FACS-based Protocol to Isolate RNA from the Secondary Cells of Drosophila Male Accessory Glands. J. Vis. Exp. (2019).
This video describes the dissection of the accessory glands from male flies and shows an example in which the isolated gland will be dissociated into a single cell suspension.
Protocol
This protocol is an excerpt from Immarigeon et al., A FACS-based Protocol to Isolate RNA from the Secondary Cells of Drosophila Male Accessory Glands, J. Vis. Exp. (2019). 1. Solutions and Material Preparation Prepare the following solutions. Prepare Serum Supplemented Medium (SSM): Schneider's Drosophila medium complemented with 10% heat inactivated fetal bovine serum and 1% Penicillin-Streptomycin.</li…
Materials
| Binocular microscope for dissection | |||
| Binocular with light source for GFP | |||
| Bunsen | |||
| Plastic microtubes 1.5 mL | Eppendorf | ||
| Fine dissection forceps | |||
| Foetal bovine serum | Gibco | 10270-106 | heat inactivated prior to use |
| Glass dishes for dissection | |||
| Ice bucket | |||
| P1000 | Gilson | ||
| P20 | Gilson | ||
| P200 | Gilson | ||
| Papain | 50U/mL stock | ||
| PBS | home made | ||
| Penicillin-Streptomycin | Gibco | 15070-063 | |
| Schneider’s Drosophila medium | Gibco | 21720-001 | |
| Tipone 1250 μL graduated tip | Starlab | S1161-1820 | |
| Tipone 200 μL bevelled tip | Starlab | S1161-1800 | |
| TrypLE Express Enzyme | Gibco | 12604013 |
Cite This Article
Drosophila Male Accessory Gland Dissection: A Method to Isolate Secondary Cells. J. Vis. Exp. (Pending Publication), e20100, doi: (2023).