Nile Red Fluorescence

Nile Red fluorescence is an optical phenomenon produced by the lipophilic dye Nile Red, which helps identify hydrophobic materials and contaminants in environmental samples. The dye exhibits weak fluorescence in water but becomes brighter and changes emission color in nonpolar environments because solvent polarity alters its excited-state behavior, allowing signal intensity and wavelength to indicate local chemical conditions. This property supports the detection and characterization of oil residues, lipid-rich particles, and microplastics in water and soil. Combined with fluorescence microscopy or spectroscopic analysis, Nile Red staining can improve contaminant screening, particle classification, and monitoring of environmental pollution.

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JoVE EoE - Staining Techniques

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

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2025

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

Nile Red Staining of C. elegans: A Method to Visualize Lipid Droplets in Fixed Animals

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2023

This video describes a protocol to stain lipids in whole, fixed worms using Nile Red dye. To specifically view the neutral lipids in the core of lipid droplets, image the green emission spectra of the stained C. elegans.

Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining

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Cited by 121 •

2018

Nile red staining of fixed Caenorhabditis elegans is a method for quantitative measurement of neutral lipid deposits, while oil red O staining facilitates qualitative assessment of lipid distribution among tissues.

Measuring Mast Cell Exocytosis via Neuropeptide Y Monomeric Red Fluorescent Protein Release

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2025

This video demonstrates mast cell exocytosis induced by an activating reagent that includes a calcium ionophore, initiating the release of secretory granules containing the fluorescent reporter protein NPY-mRFP. Subsequently, we quantitatively assess the exocytosis by measuring the released NPY-mRFP using a fluorescence plate reader.

A Simple and Rapid Protocol for Measuring Neutral Lipids in Algal Cells Using Fluorescence

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Cited by 32 •

2014

A simple protocol to determine the neutral lipid content of algal cells using a Nile Red staining procedure is described. This time-saving technique offers an alternative to traditional gravimetric-based lipid quantification protocols. It has been designed for the specific application of monitoring bioprocess performance.

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