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Cell Navigator® Fluorimetric Lipid Droplet Assay Kit *Red Fluorescence*
Lipid droplets, also referred to as lipid bodies, oil bodies or adiposomes, are lipid-rich cellular organelles that regulate the storage and hydrolysis of neutral lipids. They also serve as a reservoir of lipid source for many important biological processes such as fatty acid and cellular cholesterol for energy and membrane formation and maintenance. Abnormal accumulation of the cytoplasmic lipid droplets occurs in a variety of pathological conditions and can be an indicator of metabolic deficiency or pathogenesis. AAT Bioquest's Cell Navigator® Fluorimetric Lipid Droplet Assay Kit is a robust tool that could quantitatively measure lipid droplet accumulation. Droplite™ Red is used in the kit for lipophilic stain. Droplite™ Red is intensely fluorescent in a lipid-rich environment while it has minimal fluorescence in aqueous media. It is an excellent vital stain for the detection of intracellular lipid droplets with fluorescence microscopy, flow cytometry or fluorescence microplate reader. The red fluorescence signal could be read observed using the filter set of TRITC.
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
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.
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
Product family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Correction Factor (260 nm) | Correction Factor (280 nm) |
| Cell Navigator® Fluorimetric Lipid Droplet Assay Kit *Blue Fluorescence* | 336 | 443 | - | - | - |
| Cell Navigator® Fluorimetric Lipid Droplet Assay Kit *Green Fluorescence* | 504 | 510 | 81000 | 0.015 | 0.018 |
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)
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)
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)
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
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
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
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
Authors: Takahashi, H.; Kutasy, B.; Friedmacher, F.; Takahashi, T.; Puri, P.
Journal: Pediatr Surg Int (2016): 155-60
Page updated on December 17, 2024
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