Cell Navigator® Fluorimetric Lipid Droplet Assay Kit Red Fluorescence

Fluorescence images of intracellular lipid droplets in control (Left) and Oleic Acid treated HeLa cells (Right) using Cell Navigator® Lipid Droplets Fluorescence Assay Kit. HeLa cells were incubated with 300 uM of Oleic Acid for 24 hours to induce intracellular lipid droplets formation. After washing with PBS, the cells were labeled with 1X Droplite™ Red and Hoechst 33342  (Cat#17533).

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Spectral properties

Absorbance (nm)	560
Extinction coefficient (cm ^-1 M ^-1)	38000
Excitation (nm)	559
Emission (nm)	635
Quantum yield	0.7000¹

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12352200

Alternative formats

Cell Navigator® Fluorimetric Lipid Droplet Assay Kit *Green Fluorescence*

Overview SDS Protocol

Platform

Fluorescence microscope

Excitation	TRITC filter set
Emission	TRITC filter set
Recommended plate	Black wall/clear bottom

Fluorescence microplate reader

Excitation	550 nm
Emission	640 nm
Cutoff	610 nm
Recommended plate	Solid black

Components

Example protocol

AT A GLANCE

Protocol Summary

Prepare cells with test compounds
Add 100 µL Droplite™ Red staining solution
Incubate at room temperature or 37°C for 10 to 30 min
Read fluorescence intensity at Ex/Em = 550/640 nm (Cutoff = 610 nm), image cells using fluorescence microscope with TRITC filter or flow cytometer with FL1 channel

Important Following is our recommended protocol for live cells. This protocol only provides a guideline, and should be modified according to your specific needs. Thaw all the kit components at room temperature before starting the experiment.

CELL PREPARATION

For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

Droplite™ Red staining solution

Dilute 2 µL of Droplite™ Red (Component A) to 1 mL of Staining Buffer (Component B). Protect from light. Note: 20 µL of Droplite™ Red (Component A) is enough for one 96-well plate. The optimal concentration of the Droplite™ Red varies depending on specific applications. The staining conditions may be modified according to a particular cell type and the permeability of the cells or tissues to the probe.

PREPARATION OF WORKING SOLUTION

Add 2 µL of Droplite™ Red (Component A) to 1 mL of Staining Buffer (Component B) to make Droplite™ Red staining soluton. Protect from light.
Note 20 µL of Droplite™ Red (Component A) is enough for one 96-well plate. The optimal concentration of the Droplite™ Red varies depending on specific applications. The staining conditions may be modified according to a particular cell type and the permeability of the cells or tissues to the probe.

SAMPLE EXPERIMENTAL PROTOCOL

For adherent cells:

Grow cells either in a 96-well black wall/clear bottom plate (100 µL/well/96-well) or on cover-slips inside a petri dish filled with the appropriate culture medium.
Gently aspirate the culture medium, and add equal volume (such as 100 µL/well/96-well plate) of the Droplite™ Red staining solution.
Incubate the cells in a 37 °C, 5% CO₂ incubator for 10 - 30 minutes.
Remove Droplite™ Red staining solution (Optional).
Read the fluorescence intensity with a microplate reader at Ex/Em =550/640 nm (Cutoff = 610 nm) or observe the cells using a fluorescence microscope with a TRITC filter set.

For suspension cells:

Centrifuge the cells at 1000 rpm for 5 minutes to get 1 - 5 × 10⁵ cells per tube.
Resuspend cells in 500 µL of Droplite™ Red staining solution.
Incubate at room temperature or 37 °C for 10 to 30 min, protected from light.
Centrifuge to remove the Droplite™ Red staining solution, and resuspend cells in 500 µL of pre-warmed medium or buffer of your choice to get 1 - 5 × 10⁵ cells per tube (Optional).
Monitor the fluorescence increase using fluorescence microscope with a TRITC filter set or flow cytometer with FL1 channel.
Note Since Droplite™ Red has minimal fluorescence in aqueous media, aspiration of the growth medium and removal of Droplite™ Red staining solution after staining is optional. Stained cells can be fixed with 3 - 4% formaldehyde. In addition, prefixed cells (3 - 4% formaldehyde fixation) can be stained with Droplite™ Red staining solution.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Absorbance (nm)	560
Extinction coefficient (cm ^-1 M ^-1)	38000
Excitation (nm)	559
Emission (nm)	635
Quantum yield	0.7000¹

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Correction Factor (280 nm)
Cell Navigator® Fluorimetric Lipid Droplet Assay Kit Green Fluorescence	504	510	81000	0.018

Images

Figure 1. Fluorescence images of intracellular lipid droplets in control (Left) and Oleic Acid treated HeLa cells (Right) using Cell Navigator® Lipid Droplets Fluorescence Assay Kit. HeLa cells were incubated with 300 uM of Oleic Acid for 24 hours to induce intracellular lipid droplets formation. After washing with PBS, the cells were labeled with 1X Droplite™ Red and Hoechst 33342 (Cat#17533).

Citations

View all 2 citations: Citation Explorer

Biophysical and genetic cues regulating the structural remodeling of adipose tissue upon caloric excess
Authors: Zhou, Weinan and Bandara, Sarith and Leal, Cecilia and Anakk, Sayeepriyadarshini
Journal: (2022)

Impact of regulative noise exposure to biodiesel production due to enhanced lipid droplet production in Saccharomyces cerevisiae: Preliminary results from a laboratory experiment
Authors: Kumar, Reetesh
Journal: bioRxiv (2020)

References

View all 26 references: Citation Explorer

Live cell imaging and analysis of lipid droplets biogenesis in HCV infected cells
Authors: Nevo-Yassaf, I.; Lovelle, M.; Nahmias, Y.; Hirschberg, K.; Sklan, E. H.
Journal: Methods (2017)

Lipid droplets and associated proteins in sebocytes
Authors: Schneider, M. R.
Journal: Exp Cell Res (2016): 205-8

and beyond
Authors: Wang, C. W., Lipid droplets, lipophagy
Journal: Biochim Biophys Acta (2016): 793-805

Acute accumulation of free cholesterol induces the degradation of perilipin 2 and Rab18-dependent fusion of ER and lipid droplets in cultured human hepatocytes
Authors: Makino, A.; Hullin-Matsuda, F.; Murate, M.; Abe, M.; Tomishige, N.; Fukuda, M.; Yamashita, S.; Fujimoto, T.; Vidal, H.; Lagarde, M.; Delton, I.; Kobayashi, T.
Journal: Mol Biol Cell (2016): 3293-3304

Expression of hepatic lipid droplets is decreased in the nitrofen model of congenital diaphragmatic hernia
Authors: Takahashi, H.; Kutasy, B.; Friedmacher, F.; Takahashi, T.; Puri, P.
Journal: Pediatr Surg Int (2016): 155-60

The life cycle of lipid droplets
Authors: Hashemi, H. F.; Goodman, J. M.
Journal: Curr Opin Cell Biol (2015): 119-24

distribution and saturation level in Non-Alcoholic Fatty Liver Disease in mice
Authors: Kochan, K.; Maslak, E.; Krafft, C.; Kostogrys, R.; Chlopicki, S.; Baranska, M., Raman spectroscopy analysis of lipid droplets content
Journal: J Biophotonics (2015): 597-609

Spastin binds to lipid droplets and affects lipid metabolism
Authors: Papadopoulos, C.; Orso, G.; Mancuso, G.; Herholz, M.; Gumeni, S.; Tadepalle, N.; Jungst, C.; Tzschichholz, A.; Schauss, A.; Honing, S.; Trifunovic, A.; Daga, A.; Rugarli, E. I.
Journal: PLoS Genet (2015): e1005149

Structure, function and metabolism of hepatic and adipose tissue lipid droplets: implications in alcoholic liver disease
Authors: Natarajan, S. K.; Rasineni, K.; Ganesan, M.; Feng, D.; McVicker, B. L.; McNiven, M. A.; Osna, N. A.; Mott, J. L.; Casey, C. A.; Kharb and a, K. K.
Journal: Curr Mol Pharmacol (2015)

Chemical imaging of lipid droplets in muscle tissues using hyperspectral coherent Raman microscopy
Authors: Billecke, N.; Rago, G.; Bosma, M.; Eijkel, G.; Gemmink, A.; Leproux, P.; Huss, G.; Schrauwen, P.; Hesselink, M. K.; Bonn, M.; Parekh, S. H.
Journal: Histochem Cell Biol (2014): 263-73