Live Staining of Lipid Droplets Using Nile Red



April 7, 2019

EP149 v1.0

Goals:

Key Steps:

Other Required Materials:

Visualize lipid droplets in Organ-Chips

  • Live staining and fluorescent imaging in Organ-Chips
  • Nile red (Invitrogen™ N1142)
  • Cell culture medium
  • Fluorescence microscope

Optional:

  • PBS
  • 4% paraformaldehyde (PFA)
  • Fluorescence microscope

Background

Nile red is used to localize and quantify lipids, particularly neutral lipid droplets within cells.

Method

Sample type

Live Organ-Chip

See Protocol EP155 Live Staining of Cells.

Recommended reagent dilution and incubation time

Nile Red (Invitrogen™ N1142)

Prepare 30 mM stock in DMSO. Dilute 1:3000 in cell culture medium. Incubate for 30 minutes at room temperature.

Fixative (optional)

Chips can be fixed immediately after staining: 4% PFA for 15 minutes at room temperature.

See Protocol EP137 Fixation and Immunofluorescence (IF) Staining.

Representative image

../../../../../Desktop/Protocols/Protocols-Staining/NileRed,DAPI_3.jpg Image of Nile red staining showing lipid droplets (red) / nuclei staining (blue) indicating presence of lipid droplets in hepatocytes in the human Liver-Chip (top channel)

More information on vendor site

https://www.thermofisher.com/order/catalog/product/N1142

 


 

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