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Fluorescent Probe for Lipid Droplet Imaging: ShoyakuGreen

ShoyakuGreen (= TFMAQ-8Ph) is a green fluorescent dye (λex, max = 404 nm, λem, max = 473 nm) which not only has a high specificity for fat droplets but also low cytotoxicity.

ShoyakuGreen (= TFMAQ-8Ph)

Product

This product was commercialized under instruction of Prof. Satoru Karasawa, Dr. Yasufumi Fuchi, and Dr. Koichi Hamada.

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Advantages

  • Maximum excitation wavelength at 404 nm, Maximum emission wavelength at 473 nm (in n-hexane)
  • Strong and specific fluorescence imaging of lipid droplets
  • Low cytotoxicity

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Application: Intracellular Lipid Droplet Imaging

Fluorescent imaging of cells stained with hoyakuGreen (green)とPI (red)

Figure. Fluorescent imaging of cells stained with ShoyakuGreen (green)PI (red) (A:3T3-L1 cell、B:HeLa cell)
(Observation via Leica TCS SP5)

[Cell Pre-Treatment]

A) 3T3-L1 cells were incubated with differentiation medium in a 5% CO2 incubator at 37°C for 10 days.
B) HeLa cells were incubated with oleic acid in a 5 % CO2 incubator at 37°C for 48 hours.

[Preparation of ShoyakuGreen Solution]

  1. Stock solution: ShoyakuGreen was dissolved in 100% DMSO at a concentration of 1 mM.
  2. Working solution: The above stock solution was diluted to a final working concentration of 1 µM in culture medium.

[Fluorescence Staining of Intracellular Lipid Droplets]

  1. Remove culture medium and Replace with 1 µM ShoyakuGreen working solution (Green).
  2. Incubate cells in a 5 % CO2 incubator at 37°C for 30 minutes.
  3. Wash – PBS x 3
  4. Fixation - 4% PFA/PB, RT, 10 minutes
  5. Wash – PBS x 3
  6. Permeabilization - 0.1% Triton X-100/PBS, RT, 15 minutes
  7. Wash – PBS x 3
  8. RNase Treatment – 100 µg/mL RNase/PBS, 37°C, 20 minutes
  9. Wash – PBS x 3
  10. PI Staining – add 5 µg/mL PI/PBS (red), 15 minutes
  11. Wash – PBS x 3
  12. Observe lipid droplets using an appropriate fluorescence microscope.

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References

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Related Product Category Page

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