CytoTell™ Blue
Overview | ![]() ![]() |
Molecular weight 379.71 | Excitation (nm) 410 | Emission (nm) 445 |
Platform
Flow cytometer
Excitation | 405 nm laser |
Emission | 450/40 nm filter |
Instrument specification(s) | Pacific Blue channel |
Example protocol
AT A GLANCE
Protocol summary
- 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
Important notes
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.
Product Number |
Indicator |
Size |
Ex/Em (nm) |
Excitation Source |
22240 |
CytoTell™ UltraGreen |
500 tests |
492/519 |
488 nm (Blue Laser) |
22241 |
CytoTell™ UltraGreen |
1000 tests |
492/519 |
488 nm (Blue Laser) |
22248 |
CytoTell™ Violet 500 |
500 tests |
415/499 |
405 nm (Violet Laser) |
22251 |
CytoTell™ Blue |
500 tests |
403/454 |
405 nm (Violet Laser) |
22252 |
CytoTell™ Blue |
1000 tests |
403/454 |
405 nm (Violet Laser) |
22253 |
CytoTell™ Green |
500 tests |
511/525 |
488 nm (Blue Laser) |
22254 |
CytoTell™ Green |
1000 tests |
511/525 |
488 nm (Blue Laser) |
22255 |
CytoTell™ Red 650 |
500 tests |
628/643 |
633 nm (Red Laser) |
22256 |
CytoTell™ Red 650 |
1000 tests |
628/643 |
633 nm (Red Laser) |
22257 |
CytoTell™ Orange |
500 tests |
542 /556 |
488 nm (Blue Laser) |
22258 |
CytoTell™ Orange |
1000 tests |
542 /556 |
488 nm (Blue Laser) |
22261 |
CytoTell™ Red 590 |
500 tests |
560 /574 |
488 nm (Blue Laser) |
22262 |
CytoTell™ Red 590 |
1000 tests |
560 /574 |
488 nm (Blue Laser) |
PREPARATION OF STOCK SOLUTION
CytoTell™ dye stock solution (500X):
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
CytoTellTM dye working solution (1X):
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.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 263.359 µL | 1.317 mL | 2.634 mL | 13.168 mL | 26.336 mL |
5 mM | 52.672 µL | 263.359 µL | 526.718 µL | 2.634 mL | 5.267 mL |
10 mM | 26.336 µL | 131.679 µL | 263.359 µL | 1.317 mL | 2.634 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Citations
Authors: Jin, Liyuan and Cao, Lei and Zhu, Yingjie and Cao, Jiani and Li, Xiaoyan and Zhou, Jianxia and Liu, Bing and Zhao, Tongbiao
Journal: Science Bulletin (2020)
Authors: Huang, Jianhan and Zheng, Meijun and Zhang, Zongliang and Tang, Xin and Chen, Yaxing and Tong, Aiping and Zhou, Liangxue
Journal: (2020)
Authors: Sawada, Masaaki and Goto, Kumiko and Morimoto-Okazawa, Akiko and Haruna, Miya and Yamamoto, Kei and Yamamoto, Yoko and Nakagawa, Satoshi and Hiramatsu, Kosuke and Matsuzaki, Shinya and Kobayashi, Eiji and others,
Journal: International Immunology (2020): 397--405
Authors: Rajasinghe, Lichchavi D and Chauhan, Preeti S and Wierenga, Kathryn A and Evered, Augustus O and Harris, Shamya N and Bates, Melissa A and Gavrilin, Mikhail A and Pestka, James J
Journal: Frontiers in immunology (2020): 2179
Authors: Nakagawa, Naoki and Plestant, Charlotte and Yabuno-Nakagawa, Keiko and Li, Jingjun and Lee, Janice and Huang, Chu-Wei and Lee, Amelia and Krupa, Oleh and Adhikari, Aditi and Thompson, Suriya and others,
Journal: Neuron (2019): 836--852
Authors: Nakajima, Masao and Sakoda, Yukimi and Adachi, Keishi and Nagano, Hiroaki and Tamada, Koji
Journal: Cancer science (2019): 3079--3088
Authors: Vitali, Ilaria and Fi{\`e}vre, Sabine and Telley, Ludovic and Oberst, Polina and Bariselli, Sebastiano and Frangeul, Laura and Baumann, Natalia and McMahon, John J and Klingler, Esther and Bocchi, Riccardo and others,
Journal: Cell (2018): 1264--1276
Authors: Govindan, Subashika and Oberst, Polina and Jabaudon, Denis
Journal: Nature protocols (2018): 2297--2311
Authors: Klöss, Volker
Journal: (2017)
Authors: Klöss, Volker and Grünvogel, Oliver and Wabnitz, Guido and Eigenbrod, Tatjana and Ehrhardt, Stefanie and Lasitschka, Felix and Lohmann, Volker and Dalpke, Alex and er H, undefined
Journal: Frontiers in Immunology (2017): 1238
References
Authors: Khamesipour A, Nateghi Rostami M, Tasbihi M, Miramin Mohammadi A, Shahrestani T, Sarrafnejad A, Sohrabi Y, Esk and ari SE, Keshavarz Valian H.
Journal: Microbes Infect (2012): 702
Authors: Miao H, Jin X, Perelson AS, Wu H.
Journal: Bull Math Biol (2012): 300
Authors: Alex, undefined and er S, Sasse A, Konschalla S, Kroh A, Merx MW, Weber C, Liehn EA.
Journal: J Cell Mol Med (2012): 1640
Authors: Deleyrolle LP, Rohaus MR, Fortin JM, Reynolds BA, Azari H.
Journal: J Vis Exp. (2012)
Authors: Quah BJ, Parish CR.
Journal: J Immunol Methods (2012): 1
Authors: Newman SL, Lemen W, Smulian AG.
Journal: Med Mycol (2011): 356
Authors: Levitsky J, Gallon L, Miller J, Tambur AR, Leventhal J, Flaa C, Huang X, Sarraj B, Wang E, Mathew JM.
Journal: Transplantation (2011): 199
Authors: Wagner Z, Tabi T, Zachar G, Csillag A, Szoko E.
Journal: Electrophoresis (2011): 2816
Authors: Iverson S, Kautiainen A, Ip J, Uetrecht JP.
Journal: Chem Res Toxicol (2010): 1184
Authors: Quah BJ, Parish CR.
Journal: J Vis Exp. (2010)
FAQ
What are the most popular green DNA dyes?
What does Cytochalasin D do?
What filters should I use for Apoptosis/Necrosis Assay Kit (Cat No. 22840)?
What filters should I use for Apoptosis/Necrosis Assay Kit (Cat No. 22843)?
What size Latex Beads is used in your Cell Meter™ Fluorimetric Phagocytosis Assay Kit (Cat# 21209)?