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Nile red staining by flow cytometry

SF Shunsuke Funakoshi
IF Ian Fernandes
OM Olya Mastikhina
DW Dan Wilkinson
TT Thinh Tran
WD Wahiba Dhahri
AM Amine Mazine
DY Donghe Yang
BB Benjamin Burnett
JL Jeehoon Lee
SP Stephanie Protze
GB Gary D. Bader
SN Sara S. Nunes
ML Michael Laflamme
GK Gordon Keller
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To quantify the lipid droplets, Cayman’s Lipid Droplets Fluorescence Assay Kit (Cayman) was used. After the EBs were dissociated as described above, cells were fixed with a Fixative Solution for 10 min at room temperature. The cells were then washed with the Assay Buffer and stained with the Nile Red Staining Solution at room temperature for 15 min. The cells were again washed with the Assay Buffer and analyzed with filter sets to detect FITC using a LSR II Flow cytometer (BD PharMingen). Mean Fluorescent Intensity (MFI) was measured using FlowJo software (Tree Star). The gating strategy is shown in Supplementary Fig. 11c.

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