CytoTell™ Blue

Name: CytoTell™ Blue
Brand: AAT Bioquest
SKU: 22251
Availability: InStock

Flow cytometry combined with fluorescence staining is a powerful tool to analyze heterogeneous cell populations. Among all the existing fluorescent dyes CFSE is the preferred cell proliferation indicator that is widely used for live cell analysis. However, it is impossible to use CFSE and its fluorescein analogs for GFP-transfected cells or for the applications where a FITC-labeled antibody is used since CFSE and its fluorescein analogs have the excitation and emission spectra almost identical to GFP or FITC. CytoTell™ dyes are well excited at major laser lines such as 405 nm, 488 nm or 633 nm with multicolor emissions. CytoTell™ dyes have minimal cytotoxicity, and are used for the multicolor applications with either GFP cell lines or FITC-labeled antibodies since they have either excitation or emission spectra distinct from fluorescein. CytoTell™ Blue is a blue fluorescent dye that stains cells evenly. As cells divide, the dye is distributed equally between daughter cells that can be measured as successive halving of the fluorescence intensity of the dye. Cells labeled with CytoTell™ Blue may be fixed and permeabilized for analysis of intracellular targets using standard formaldehyde-containing fixatives and saponin-based permeabilization buffers. CytoTell™ Blue has a peak excitation of 405 nm and can be excited by the violet (405 nm) laser line. It has a peak emission of 450 nm and can be detected with a 450/20 band pass filter (equivalent to Pacific Blue®, or BD Horizon® V450), making it compatible with applications that utilize GFP or FITC antibodies for multicolor cell analysis.

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) 531 nm (Green Laser)
22258	CytoTell™ Orange	1000 tests	542 /556	488 nm (Blue Laser) 531 nm (Green Laser)
22261	CytoTell™ Red 590	500 tests	560 /574	488 nm (Blue Laser) 531 nm (Green Laser)
22262	CytoTell™ Red 590	1000 tests	560 /574	488 nm (Blue Laser) 531 nm (Green Laser)

PREPARATION OF STOCK SOLUTION

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.

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

CytoTell^TM 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

Table 1. Volume of DMSO needed to reconstitute specific mass of CytoTell™ Blue to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	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

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Spectrum

Open in Advanced Spectrum Viewer

Product family

Name	Excitation (nm)	Emission (nm)
CytoTell™ Green	510	525
CytoTell™ Orange	541	560

Citations

View all 19 citations: Citation Explorer

Sustained inhibition of CSF1R signaling augments antitumor immunity through inhibiting tumor-associated macrophages
Authors: Sato, Takahiko and Sugiyama, Daisuke and Koseki, Jun and Kojima, Yasuhiro and Hattori, Satomi and Sone, Kazuki and Nishinakamura, Hitomi and Ishikawa, Tomohiro and Ishikawa, Yuichi and Kato, Takuma and others,
Journal: JCI insight (2025): e178146

Inhibition of furin in CAR macrophages directs them toward a proinflammatory phenotype and enhances their antitumor activities
Authors: Ziane-Chaouche, Lydia and Raffo-Romero, Antonella and Hajjaji, Nawale and Kobeissy, Firas and Pinheiro, Donna and Aboulouard, Soulaimane and Cozzani, Adeline and Mitra, Suman and Fournier, Isabelle and Cizkova, Dasa and others,
Journal: Cell Death \& Disease (2024): 1--16

The CH1 domain influences the expression and antigen sensing of the HIV-specific CH31 IgM-BCR and IgG-BCR
Authors: Ortiz, Yaneth and Anasti, Kara and Pane, Advaiti K and Cronin, Kenneth and Alam, S Munir and Reth, Michael
Journal: Proceedings of the National Academy of Sciences (2024): e2404728121

Vaccine plus microbicide effective in preventing vaginal SIV transmission in macaques
Authors: Rahman, MA and Bissa, M and Silva de Castro, I and Helmold Hait, S and Stamos, JD and Bhuyan, F and Hunegnaw, R and Sarkis, S and Gutowska, A and Doster, MN and others,
Journal: Nature Microbiology (2023): 1--14

The impact of CCR8+ regulatory T cells on cytotoxic T cell function in human lung cancer
Authors: Haruna, Miya and Ueyama, Azumi and Yamamoto, Yoko and Hirata, Michinari and Goto, Kumiko and Yoshida, Hiroshi and Higuchi, Naoko and Yoshida, Tetsuya and Kidani, Yujiro and Nakamura, Yamami and others,
Journal: Scientific Reports (2022): 1--12

References

View all 50 references: Citation Explorer

Phenotyping of circulating CD8(+) T cell subsets in human cutaneous leishmaniasis
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

Evaluation of multitype mathematical models for CFSE-labeling experiment data
Authors: Miao H, Jin X, Perelson AS, Wu H.
Journal: Bull Math Biol (2012): 300

Repetitive transplantation of different cell types sequentially improves heart function after infarction
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

Identification and isolation of slow-dividing cells in human glioblastoma using carboxy fluorescein succinimidyl ester (CFSE)
Authors: Deleyrolle LP, Rohaus MR, Fortin JM, Reynolds BA, Azari H.
Journal: J Vis Exp. (2012)

New and improved methods for measuring lymphocyte proliferation in vitro and in vivo using CFSE-like fluorescent dyes
Authors: Quah BJ, Parish CR.
Journal: J Immunol Methods (2012): 1

Page updated on July 30, 2025

Ordering information

Price
Unit size	500 Tests 2x500 Tests
Catalog Number	2225122252
Quantity

Add to cart

Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Technical Support	Contact us
Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Request quotation

Physical properties

Molecular weight	379.71
Solvent	DMSO

Spectral properties

Excitation (nm)	410
Emission (nm)	445

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200