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Droplite™ Green

Lipid Droplets (LDs) are essential organelles responsible for storing neutral lipids, primarily consisting of triglycerides and cholesterol esters. Found in a variety of cell types, LDs play crucial roles in numerous biological processes, including metabolism, membrane biosynthesis, cell signaling, inflammation, and cancer-related mechanisms. Droplite™ Green, developed by AAT Bioquest, is a green fluorescent dye with an extremely high affinity for LDs and minimal cell toxicity. This probe has an excitation and emission maxima of 421 nm and 521 nm, respectively, and can be conveniently detected using fluorescence microscopy or an HCS reader. In cell culture experiments, Droplite™ Green demonstrates exceptional safety, even at high concentrations, ensuring reliable and accurate LD characterization without compromising cell health and viability.

Example protocol

AT A GLANCE

Protocol Summary
  1. Prepare and treat cells in a growth medium.
  2. Incubate cells with Droplite™ Green working solution for 20 to 30 minutes at 37 °C.
  3. Remove Droplite™ Green working solution.
  4. Add HHBS buffer and analyze using a fluorescence microscope equipped with a FITC filter set.
Important

The 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 adherent cells
  1. Plate cells overnight in growth medium at 10,000 to 40,000 cells/well/90 μL for a 96-well plate or 2,500 to 10,000 cells/well/20 μL for a 384-well plate.

For non-adherent cells
  1. Centrifuge the cells from the culture medium.

  2. Suspend the cell pellets in culture medium at 50,000-100,000 cells/well/90 µL for a 96-well poly-D lysine plate or 10,000-25,000 cells/well/20 µL for a 384- well poly-D lysine plate.

  3. Centrifuge the plate at 800 rpm for 2 minutes with the brake off prior to your experiment.

    Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density.

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™ Green stock solution
  1. Add 100 µL of DMSO into the Droplite™ Green vial and mix well.

    Note: 100 µL stock solution is enough for 100 tests. The staining conditions may be modified according to the particular cell type.

    Note: Make a single unused Droplite™ Green stock solution aliquot and store it at ≤ -20 °C. Protect from light and avoid repeated freeze-thaw cycles.

PREPARATION OF WORKING SOLUTION

Droplite™ Green working solution
  1. Add 100 µL of Droplite™ Green stock solution into 10 mL of a buffer of your choice or cell culture medium, and mix well.

    Note: HHBS [Hanks' Buffer with 20 mM Hepes] buffer (AAT Cat# 20011) can be used to make a working solution. Prepare a fresh working solution just before use.

SAMPLE EXPERIMENTAL PROTOCOL

Stain cells
  1. Prepare and treat cells in a growth medium as desired.

  2. Add 100 µL/well (96-well plate) of Droplite™ Green working solution to the cell plate.

    Note: The optimal concentration of the cell membrane probe varies depending on the specific application.

  3. Incubate the cells at 37 °C for 20 to 30 minutes, protected from light.

  4. Remove the working solution in each well.

  5. Wash twice with DPBS and add HHBS or DPBS solution to the wells.

  6. Observe the fluorescence signal in cells using a fluorescence microscope equipped with a FITC filter set.

Spectrum

References

View all 50 references: Citation Explorer
One amino acid drives the lipid droplet targeting sequence of a new noncoding RNA-encoded protein to mitochondrion.
Authors: Yuan, Quan and Zheng, Kaixuan and Huang, Ting and Zhi, Zelun and Xu, Yilu and Cui, Liujuan and Liu, Pingsheng and Zhang, Shuyan
Journal: Proteomics (2023): e2200301
Constructing lipid droplet-targeting photosensitizers based on coumarins with NIR emission.
Authors: Guo, Yimin and Liu, Weimin and Sha, Jie and Li, Xuewei and Ren, Haohui and Wu, Jiasheng and Zhang, Wenjun and Lee, Chun-Sing and Wang, Pengfei
Journal: Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy (2023): 122698
Peroxynitrite/Lipid Droplet Sequence-Activated Dual-Lock Fluorescent Probes Enable Precise Intraoperative Imaging of Atherosclerotic Plaques.
Authors: Sang, Mangmang and Huang, Yibo and Liu, Zhiwei and Li, Gang and Wang, Yan and Yuan, Zhenwei and Dai, Cuilian and Zheng, Jinrong
Journal: ACS sensors (2023): 893-903
Bsc2 is a novel regulator of triglyceride lipolysis that demarcates a lipid droplet subpopulation.
Authors: Speer, Natalie Ortiz and Braun, R Jay and Reynolds, Emma Grace and Swanson, Jessica M J and Henne, W Mike
Journal: bioRxiv : the preprint server for biology (2023)
Two-photon excited red-green "discoloration" bioprobes for monitoring lipid droplets and lipid droplet-lysosomal autophagy.
Authors: Liu, Ming-Xuan and Xu, Li and Zhu, Peng-Fei and Li, Xin and Shan, Miao and Jin, Wei and Chen, Jing and Ling, Yong and Zhang, Xiao-Ling
Journal: Journal of materials chemistry. B (2023): 3186-3194
Page updated on October 8, 2024

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Catalog Number22729
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Physical properties

Molecular weight

432.37

Solvent

DMSO

Spectral properties

Excitation (nm)

421

Emission (nm)

521

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

Platform

Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall, clear bottom
Instrument specification(s)FITC filter set
Fluorescence images of intracellular lipid droplets in control (left) and Oleic Acid treated HeLa cells (right) using Droplite™ Green. HeLa cells were incubated with 100 uM of Oleic Acid for 19 hours to induce intracellular lipid droplet formation. After washing with DPBS, HHBS was added to the cells, and images were acquired with a fluorescence microscope using a FITC filter set.
Fluorescence images of intracellular lipid droplets in control (left) and Oleic Acid treated HeLa cells (right) using Droplite™ Green. HeLa cells were incubated with 100 uM of Oleic Acid for 19 hours to induce intracellular lipid droplet formation. After washing with DPBS, HHBS was added to the cells, and images were acquired with a fluorescence microscope using a FITC filter set.
Fluorescence images of intracellular lipid droplets in control (left) and Oleic Acid treated HeLa cells (right) using Droplite™ Green. HeLa cells were incubated with 100 uM of Oleic Acid for 19 hours to induce intracellular lipid droplet formation. After washing with DPBS, HHBS was added to the cells, and images were acquired with a fluorescence microscope using a FITC filter set.
Fluorescence imaging of intracellular lipid droplets in oleic acid-treated HeLa cells using Droplite™ Green, both before and after fixation. HeLa cells were treated with 100 µM oleic acid for 19 hours to induce intracellular lipid droplet formation. Following treatment, the cells were washed with DPBS and incubated in HHBS. Fluorescence images were captured using a FITC filter set on a fluorescence microscope. The cells were then fixed with 4% formaldehyde for 20 minutes at room temperature. After another DPBS wash, HHBS was added again, and fluorescence images were taken using the same microscope settings and exposure time.