Embryo lipid staining was performed as described previously [33 (link)]. In short, embryos were obtained by alkaline hypochlorite treatment of adult hermaphrodites, transferred to PCR tubes (Eppendorf, Hamburg, Germany) containing 4% formaldehyde in M9 buffer and subjected to 3 freeze-thaw cycles in liquid nitrogen and a 37°C water bath. Embryos were then incubated with 1 μg/ml BODIPY 493/503 (Thermo Fisher Scientific) in M9 buffer for 1 hour and washed 3 times with M9 + 0.01% Triton X-100 (Sigma #9001-93-1). Next, nuclei were stained with 5 ng/mL DAPI and imaged by epifluorescence microscopy.
Adults were fixed for lipid staining by incubating in 4% paraformaldehyde (Sigma-Aldrich 158127) for 1 hour at room temperature. Adults were then washed twice with PBS, resuspended in 95% ethanol and washed again in PBS. Next, fixed animals were incubated with 1 μg/ml BODIPY 493/503 (Thermo Fisher Scientific) in M9 buffer for 1 hour and washed 3 times with M9 + 0.01% Triton X-100. Animals were stained with 5 ng/mL DAPI and imaged by epifluorescence microscopy.
Adults were fixed for lipid staining by incubating in 4% paraformaldehyde (Sigma-Aldrich 158127) for 1 hour at room temperature. Adults were then washed twice with PBS, resuspended in 95% ethanol and washed again in PBS. Next, fixed animals were incubated with 1 μg/ml BODIPY 493/503 (Thermo Fisher Scientific) in M9 buffer for 1 hour and washed 3 times with M9 + 0.01% Triton X-100. Animals were stained with 5 ng/mL DAPI and imaged by epifluorescence microscopy.
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