Calculator
CytoTell™ UltraGreen
Example protocol
AT A GLANCE
- Prepare cells with test compounds
- Add 1X dye working solution
- Incubate dyes with cells at room temperature or 37 oC for 10 to 30 minutes
- Remove the dye working solution
- Analyse with flow cytometer with appropriate filter set
Bring all the kit components at room temperature before starting the experiment.
Note: The CytoTell™ dyes are lyophilized powders. They should be stable for at least 6 months if store at -20 °C, protecting from light, and avoiding freeze/thaw cycles.
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
Add 500 µL DMSO into the dye powder vial, mix it well by vortexing to have a stock solution (500X).
Note: The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at < - 20 oC. Avoid repeated freeze-thaw cycles, and protect from light.
PREPARATION OF WORKING SOLUTION
Dilute the 500X DMSO stock solution at 1 to 500 in Hanks and 20 mM Hepes buffer (HHBS) or the buffer of your choice, pH 7 (such as 1 µL of 500X DMSO stock solution to 500 µL buffer) right before use. Mix them well by vortexing.
Note: The final concentration of the dye working solution should be empirically determined for different cell types and/or experimental conditions. It is recommended to test at the concentrations that are at least over ten fold range. Such as CytoTell™ Red might use much less amount in some cell types than the recommend concentrations.
SAMPLE EXPERIMENTAL PROTOCOL
- Treat cells with test compounds for a desired period of time.
- Centrifuge the cells to get 1-5 × 105 cells per tube.
Resuspend cells in 500 µL of the CytoTell™ dye working solution.
Optional: One can add the 500X DMSO stock solution into the cells directly without medium removing (such as, add 1 µL500X DMSO stock solution into 500 µL cells)
- Incubate cells with a dye solution at room temperature or 37 °C for 10 to 30 minutes, protected from light.
- Remove the dye working solution from the cells, wash the cells with HHBS or buffer of your choice. Resuspend cells in 500 µL of pre-warmed HHBS or medium to get 1-5 × 105 cells per tube.
- Monitor the fluorescence change at respected Ex/Em (see Table 1) with a flow cytometer or a fluorescence microscope.
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) |
| CytoTell™ Blue | 410 | 445 |
| CytoTell™ Green | 510 | 525 |
| CytoTell™ Orange | 541 | 560 |
Citations
Authors: Kawasaki, Kento and Noma, Kazuhiro and Kato, Takuya and Ohara, Toshiaki and Tanabe, Shunsuke and Takeda, Yasushige and Matsumoto, Hijiri and Nishimura, Seitaro and Kunitomo, Tomoyoshi and Akai, Masaaki and others,
Journal: (2023)
Authors: Kido, Jun-ichi and Hiroshima, Yuka and Kido, Rie and Yoshida, Kaya and Inagaki, Yuji and Naruishi, Koji and Kajimoto, Kazuaki and Kataoka, Masatoshi and Shinohara, Yasuo and Yumoto, Hiromichi
Journal: Journal of Periodontal Research (2022)
Authors: Shirakashi, Mirei and Maruya, Mikako and Hirota, Keiji and Tsuruyama, Tatsuaki and Matsuo, Takashi and Watanabe, Ryu and Murata, Koichi and Tanaka, Masao and Ito, Hiromu and Yoshifuji, Hajime and others,
Journal: Arthritis \& Rheumatology (2021)
Authors: Yoshida, Kayo and Yoshida, Kaya and Fujiwara, Natsumi and Seyama, Mariko and Ono, Kisho and Kawai, Hotaka and Guo, Jiajie and Wang, Ziyi and Weng, Yao and Yu, Yaqiong and others,
Journal: Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease (2021): 166236
Authors: Li, Young and Li, Kun and Zhu, Lianbang and Li, Bin and Zong, Dandan and Cai, Pengfei and Jiang, Chen and Du, Pengcheng and Lin, Jun and Qu, Kun
Journal: Genome medicine (2021): 1--18
Safety data sheet