What does Nile Red stain do?
In the assay, Nile Red dye is added to the cell samples and the dye selectively stains lipid droplets within the cells. The dye is minimally fluorescent in solution in water so it is not necessary to wash the cells or remove the dye before analysis.
What is Nile Red dye?
Nile red is a hydrophobic and metachromatic dye with poor solubility and fluorescence in water, with colour emission varying from deep red to strong yellow gold in hydrophobic environments. Depending on excitation and emission wavelength, the dye has been used to stain different hydrophobic molecules.
Is Nile Red fluorescent?
By contrast, Nile Red (NR) is a fluorescent dye that is well situated to report changes in the chemical polarity of cell membranes and myelin, being both lipophilic (23, 24) and differentially fluorescent depending on solvent environment (i.e., exhibits solvatochromism) (25).
What does bodipy measure?
BODIPY® 581/591 undecanoic acid can be used to detect reactive oxygen species (ROS) in cells and membranes. Oxidation of the polyunsaturated butadienyl portion of the dye results in a shift of the fluorescence emission peak from ∼590 nm to ∼510 nm.
What ethnicity is Nile red?
He is of German and Japanese descent. His icons are variations of the Belousov–Zhabotinsky reaction.
Is Nile red a real chemist?
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.
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.
Is Nile red fixable?
Nile dyes have strong affinity with lipids, no matter if they are in living cells, dead cells, mayonnaise or yoghurt. So you can use without any problem those dyes on fixed cells.
What is lysochrome dye?
A lysochrome is a soluble dye used for histochemical staining of lipids, which include triglycerides, fatty acids, and lipoproteins. Lysochromes such as Sudan IV dissolve in the lipid and show up as colored regions.
How do you make a red Nile solution?
The protocol is simple and rapid: Nile red is diluted from a stock solution (0.4 mg/ml in DMSO) 200-fold into water to 2 μg/ml, and a 10-fold volume excess (e.g., 50 ml staining solution for a 5 ml minigel) is added to a gel with immediate and thorough agitation.
What’s the difference between NileRed and Nileblue?
Structures of Nile red and Nile blue. Nile red has no formal charges and is intensely fluorescent with a high quantum yield in apolar media. Nile blue is a cationic dye and is thus more readily soluble in water than Nile red. It is highly sensitive to pH changes making it a useful pH probe.
What ethnicity is NileRed?
How do you stain yeast cells with Nile red?
Rostron and Lawrence (2017)proposed a detailed methodology to stain yeast cells with Nile red, however, they suggested fluorescence reading immediately after Nile red addition in the solution and recommended good practices in pipetting and mixing the yeast suspension to reduce variability in the measurements.
Can Nile red fluorescence be used to quantify neutral lipids in yeast?
Discussion Nile red fluorescence is widely used to screen new oleaginous yeasts (Hicks et al., 2019; Sitepu et al., 2019; Zhao et al., 2019), but also to quantify neutral lipids in non-oleaginous yeasts, such as S. cerevisiaeor Schizosaccharomyces pombe(Rostron et al., 2015).
Why can’t we stain lipid droplets with Nile red?
Several authors report failure of efficient cell staining with Nile red, caused by the difficulty of penetrating the cell wall, resulting in a poor fluorescence signal [ 21, 39, 84, 90, 93, 94 ]. Some studies aimed to develop new protocols that improve dye penetration and ultimate staining of lipid droplets.
What is the fluorescence of Nile red?
Nile red is used to localize and quantitate lipids, particularly neutral lipid droplets within cells. Nile red is almost nonfluorescent in water and other polar solvents but undergoes fluorescence enhancement and large absorption and emission blue shifts in nonpolar environments (excitation/emission maxima ~552/636 nm in methanol).