How does Nile Red bind to lipids?

For instance, using the excitation/emission wavelengths 450 to 500/>528, Nile red can dye the cholesterol in the human plasma through staining of lipid vesicles in smooth muscle cells and in cultured macrophages incubated at low density [33,34].

What does Nile Red stain do?

Nile red was initially defined as a highly specific vital stain, allowing the detection of intracellular LDs in cultured aortic smooth muscle cells and peritoneal macrophages, as it selectively dissolves in the lipids and fluoresces with no local interaction with the tissue constituent (Greenspan et al., 1985).

How do you detect lipid droplets?

Lipid droplets can be visualized either by staining the lipid ester core using fluorescent dyes or by labeling lipid droplet-specific proteins using antibodies. The intracellular distribution of lipid droplets can be analyzed without much difficulty by these methods, but care must be taken to avoid certain pitfalls.

Is Nile Red soluble in water?

In water, the Nile Red derivative 1 showed hardly any solubility at all. Surprisingly, despite having a phenolic-OH, this compound was still insoluble in pH 9 borate buffer (1 M) or in sodium carbonate solution. Consequently, spectroscopic data for this compound were only recorded in EtOH (Table 1).

Is Nile Red and Nile blue the same person?

Nile red (also known as Nile blue oxazone) is a lipophilic stain. Nile red stains intracellular lipid droplets yellow. In contrast, in phospholipids (polar lipids), Nile red has an excitation maximum of about 554 nm (green), and an emission maximum of about 638 nm (red).

Is Nile Red toxic?

TOXICITY AND IRRITATION NILE RED: No significant acute toxicological data identified in literature search. All waste must be handled in accordance with local, state and federal regulations. In some areas, certain wastes must be tracked.

How does Nile stain red?

We outline a five-step method to customize the Nile Red protocol to a specific strain: 1) Evaluate the strain’s suitability by checking for the presence of neutral lipid, 2) Select of the best excitation/emission wavelength, 3) Optimization of incubation time, stain carrier, dye concentration, and temperature, 4) …

How do you isolate lipid droplets?

The isolation of lipid droplets by centrifugation is a relatively simple procedure. A very low protein-to-lipid ratio renders lipid droplets more buoyant than all other subcellular structures; lipid droplets can be separated from more dense subcellular compartments using discontinuous density gradients.

What is bodipy staining?

Description. With its nonpolar structure and long-wavelength absorption and fluorescence, BODIPY® 493/503 can be used as a stain for neutral lipds and as a tracer for oil and other nonpolar lipids.

How do you dissolve Red Nile?

o Nile Red Stock Solution (1 mg/mL) – dissolve 25 mg of Nile Red (Invitrogen N-1142) in 25 mL of DMSO; Store at room temperature in the dark indefinitely.

Who runs Nile red?

Nigel Braun
Nigel Braun began posting his chemistry experiments to YouTube in 2014 under the name NileRed (after the lipophilic stain) with no intention of amassing such a large audience. But now, four years later, he has over 420,000 subscribers that tune in regularly to watch his latest experiment.

Does Nile red have a degree?

Academic Background After finishing his degree, he worked as a lab tech in an organic lab and took post-graduate studies in applied chemistry. He however ended up leaving his post-graduate studies early to focus on his YouTube channel.

How is Nile red used as a lipid stain?

The dye is present as a minor component of commercial preparations of the nonfluorescent lipid stain nile blue (5). Nile red is intensely fluorescent and, if proper spectral conditions are chosen, it can serve as a sensitive vital stain for the detection of cytoplasmic lipid droplets.

How does Nile red staining kit ab228553 work?

Nile Red Staining Kit ab228553 is designed for the quantitative, fluorometric detection and measurement of intracellular lipid droplets using fluorescence microscopy, flow cytometry or a fluorescence microplate reader. In the assay, Nile Red dye is added to the cell samples and the dye selectively stains lipid droplets within the cells.

What happens when Nile red is applied to a cell?

Nile red can be applied to cells in an aqueous medium, and it does not dissolve the lipids it is supposed to reveal. The formation of cytoplasmic lipid droplets is a normal cellular process. The droplets are neutral lipids, usually tria- cylglycerols or cholesteryl esters.

What are the properties of Nile red dye?

Nile red exhibits properties of a near-ideal lysochrome. It is strongly fluorescent, but only in the presence of a hydrophobic environment. The dye is very soluble in the lipids it is intended to show, and it does not interact with any tissue constituent except by solution.