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

Published: April 30, 2023

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…

Declarações

The authors have nothing to disclose.

Materials

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

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Nile Red Staining: A Technique to Detect and Quantify Intracellular Neutral Lipids in Algae. J. Vis. Exp. (Pending Publication), e21032, doi: (2023).

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