Source: Wei, C., et al., Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae. J. Vis. Exp. (2019).
This video demonstrates lipid droplet staining in Drosophila larval oenocytes – hepatocyte-like cells – using the lipophilic fluorescent BODIPY based dye. These cells act as a model for studying lipid metabolism during development and in response to environmental stress such as starvation.
1. Starvation Treatment for the Larvae
NOTE: In the third instar larvae, there are few or no detectable lipid droplets in the oenocytes under normal feeding conditions, but numerous large lipid droplets can be induced in the oenocytes under stress conditions, such as starvation. To further verify this method, it is necessary to pretreat these larvae to induce lipid droplet biogenesis in oenocytes. Here, a starvation time course of 12 h, 24 h, and 36 h was chosen as the paradigm. In particular, a short period of starvation (e.g., 3 h) is sufficient to induce detectable lipid droplets in oenocytes. Starvation duration may vary according to specific experimental goals and settings.
2. Dissection of Oenocytes
3. Lipid Droplet Staining
4. Mounting and Imaging
The authors have nothing to disclose.
50 mL centrifuge tube | Corning | 430829 | 50 mL |
6 cm Petri dish | Thermo Fisher | 150326 | 6 cm |
Agar | For fly food | ||
Aluminum foil | N/A | N/A | Protect smaple from light |
BODIPY 493/503 | Invitrogen | D3922 | Lipid droplet staining dye |
Confocal microscope | Leica | Leica TSC SP5 | Confocal imaging |
Corn syrup | For fly food | ||
Cornmeal | For fly food | ||
Coverslip | Citoglas | 10212424C | 20 × 20 mm, 0.13-0.17 thick |
Dissection pin | N/A | N/A | |
Dissection plate | N/A | N/A | |
Filter paper | N/A | N/A | Diameter: 11 cm |
Fixation buffer | N/A | N/A | 4% Paraformaldehyde (PFA) in 1x PBS |
Forcep | Dumont | 11252-30 | #5 |
Incubator | Jiangnan | SPX-380 | For fly culture |
Microcentrifuge tube | Axygen | MCT-150-C | 1.5 mL |
Microscopy slide | Citoglas | 10127105P-G | |
Mounting medium | VECTASHIELDAntifade Mounting Medium |
H-1000 | Antifade mounting medium |
Nail polish | PanEra | AAPR419 | Seal the coverslip |
Paintbrush | N/A | N/A | |
PBS | N/A | N/A | 1x PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na_{2} HPO_{4} ,1.8 mM KH_{2} PO_{4} , pH 7.4) |
Rotator | Kylin-Bell Lab Instruments | WH-986 | |
Scissor | Smartdata Medical | SR81 | Vannas spring scissor |
Soy flour | For fly food | ||
Spatula | N/A | N/A | |
Standard cornmeal food | N/A | N/A | Accoding to Bloomington standard cornmeal food recipe |
Stereo microscope | Leica | Leica S6E | For tissue dissection |
Wipe paper | N/A | N/A | |
Yeast | For fly food |