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).