Published TCIMAIL newest issue No.197
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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.
Product
This product was commercialized under instruction of Prof. Satoru Karasawa, Dr. Yasufumi Fuchi, and Dr. Koichi Hamada.
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
Application: Intracellular Lipid Droplet Imaging
Figure. Fluorescent imaging of cells stained with ShoyakuGreen (green)と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]
- Stock solution: ShoyakuGreen was dissolved in 100% DMSO at a concentration of 1 mM.
- 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]
- Remove culture medium and Replace with 1 µM ShoyakuGreen working solution (Green).
- Incubate cells in a 5 % CO2 incubator at 37°C for 30 minutes.
- Wash – PBS x 3
- Fixation - 4% PFA/PB, RT, 10 minutes
- Wash – PBS x 3
- Permeabilization - 0.1% Triton X-100/PBS, RT, 15 minutes
- Wash – PBS x 3
- RNase Treatment – 100 µg/mL RNase/PBS, 37°C, 20 minutes
- Wash – PBS x 3
- PI Staining – add 5 µg/mL PI/PBS (red), 15 minutes
- Wash – PBS x 3
- Observe lipid droplets using an appropriate fluorescence microscope.
References
- Selective synthesis of substituted amino-quinoline derivatives by C-H activation and fluorescence evaluation of their lipophilicity-responsive properties
- Photophysical properties of emissive pyrido[3,2-c]carbazole derivatives and apoptosis induction: Development towards theranostic agents in response to light stimulus
