Nile Red Staining: A Technique to Detect and Quantify Intracellular Neutral Lipids in Algae

Abstract

Source: Storms, Z. J. et al. A Simple and Rapid Protocol for Measuring Neutral Lipids in Algal Cells Using Fluorescence. J. Vis. Exp. (2014)

This video demonstrates a protocol for detecting and quantifying neutral lipids in algal cells by staining them with Nile red dye.

Protocol

1. Fluorometric Quantification of Neutral Lipids Using Nile Red

NOTE: Only 10 µl of an algal suspension at 5 g/L is needed for the fluorescence reading. Generally, isolation of dry algal biomass from 1.5 ml of culture broth is more than sufficient. Also, the light intensity of the lamp in the spectrophotometer can degrade over time. It is recommended to include standards in every experiment to ensure that variations in the instrument do not add unnecessary error to the measurements.

1. Prepare a Nile Red solution at a concentration of 10 µg/ml dissolved in alcohol reagent grade ethanol. ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
25 ml disposable pipettes	Fisher	13-676-10K
Pipette Bulb	Fisher	13-681-51
1.5 ml microcentrifuge tubes	Fisher	05-408-129
40 ml Centrifugation tubes (FEP)	Fisher	05-562-16A	Could also use glass tubes
Pasteur glass pipettes	Fisher	13-678-20C
Nile Red	Sigma	N3013-100MG
Ethanol (alcohol reagent grade)	Fisher	AC65109-0020
Leica DMRXA2 (or equivalent) microscope	Leica	DMRXA2
Fluorescence multi-well plate reader	Thermo Lab Systems	Fluoroskan Ascen
Fluorescence reader software	Thermo Lab Systems	Ascent Software 2.6
COSTAR 96 well plate with round bottom	Fisher	06-443-2