Lipid Droplet Staining of Larval Drosophila Oenocytes: A Technique for Assessment of Lipid Droplet Accumulation Under Normal and Stressed Condition Using BODIPY-Based Dye

Published: April 30, 2023

Abstract

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.

Protocol

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 c…

Açıklamalar

The authors have nothing to disclose.

Materials

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

Etiketler

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Bu Makaleden Alıntı Yapın
Lipid Droplet Staining of Larval Drosophila Oenocytes: A Technique for Assessment of Lipid Droplet Accumulation Under Normal and Stressed Condition Using BODIPY-Based Dye. J. Vis. Exp. (Pending Publication), e21044, doi: (2023).

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