Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis of PVP
2.2. X-ray Crystallographic Data
2.3. Optical Properties
2.4. Staining Investigation on B16-F10 Melanoma Cells
2.4.1. Cell Viability
2.4.2. Fluorescence Imaging
3. Experimental Section
3.1. Materials and Methods
3.2. Synthesis
3.3. X-ray Diffraction
3.4. Fluorescence Measurements
3.4.1. One-Photon Excited Fluorescence on Solid Samples
3.4.2. Fluorescence Quantum Yield
3.5. Cell Culture and Staining Protocol
3.6. Viability Assay
3.7. Fluorescence Imaging
3.8. Lipophilicity Protocol
- (1)
- Solvent preparation (water saturated with octanol and octanol saturated with water): potassium phosphate aq buffer (0.1 M pH 7.0) and an equal volume of 1-octanol were agitated to allow saturation, then left to separate overnight.
- (2)
- Preparation of dye stock solution: 100 μg / mL in DMSO.
- (3)
- A total of 150 μL of dye stock solution was added to 1425 μL of water (Vw) and the absorbance of this initial aqueous solution was measured (A).
- (4)
- To the aqueous solution with the dye 1425 μL of 1-octanol (V0) was added.
- (5)
- The sample was stirred for 2 h, then centrifuged (3500 rpm, 10 min) to separate the phases.
- (6)
- The absorbance of the separate aqueous solution (A’) was measured.
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Cpd. | λabs a | λem b | Stokes Shift | |||||
---|---|---|---|---|---|---|---|---|
R | R′ | R″ | X− | (nm) | (nm) | (cm−1) | Ref. | |
3a | H | –CH3 | –CH3 | I− | 455 | 749 | 8627 | [11] |
3a′ | H | –CH3 | –CH3 | BF4− | 455 | 720 (605 *) | 8089 | |
3b | Br | –C2H5 | –CH3 | I− | 460 | 650 | 6354 | |
3c | H | –CH3 | –C4H9 | I− | 458 | 669 | 6886 | [11] |
3d | H | –CH3 | –(CH2)3-Cl | Br− | 459 | 670 | 6861 |
Identification Code | Dye 3b |
---|---|
Empirical formula | C22H20BrIN2S |
Formula weight | 551.27 |
Temperature/K | 123.1 |
Crystal system | Monoclinic |
Space group | P21/c |
a/Å | 17.6219(4) |
b/Å | 7.8451(2) |
c/Å | 15.0465(3) |
α/° | 90 |
β/° | 101.306(2) |
γ/° | 90 |
Volume/Å3 | 2039.75(8) |
Z | 4 |
ρcalcg/cm3 | 1.795 |
μ/mm−1 | 15.650 |
F (000) | 1080.0 |
Crystal size/mm3 | 0.1 × 0.15 × 0.17 |
Radiation | CuKα (λ = 1.54184) |
2Θ range for data collection/° | 10.238 to 141.238 |
Index ranges | −21 ≤ h ≤ 19, −9 ≤ k ≤ 5, −18 ≤ l ≤ 18 |
Reflections collected | 7087 |
Independent reflections | 3843 [Rint = 0.0299, Rsigma = 0.0447] |
Data/restraints/parameters | 3843/0/246 |
Goodness-of-fit on F2 | 1.046 |
Final R indexes [I > = 2σ (I)] | R1 = 0.0372, wR2 = 0.0961 |
Final R indexes [all data] | R1 = 0.0453, wR2 = 0.1033 |
Largest diff. peak/hole/e Å−3 | 1.11/−1.06 |
Item | Samples | Expression | Calculated/ Reported Value |
---|---|---|---|
Quantum yield | Rhodamine 101 (Ethanol + 0.01% HCl) | Φst | 1 [26] |
3a′ (Chloroform sol.) | Φx | 0.035 | |
Absorbance at 525 nm | Rhodamine 101 | Ast | 0.0265 |
3a′ | Ax | 0.07 | |
Area of emission spectrum | Rhodamine 101 | Fst | 19327.82 |
3a′ | Fx | 16255.18 | |
Average refractive index value of solvent | Rhodamine 101 | nst | 1.85423 |
3a′ | nx | 2.085 | |
Dilution ratio | Rhodamine 101 | Dst | 102 |
3a′ | Dx | 103 |
Dye | A | A′ | Log P |
---|---|---|---|
3b | 0.535 | 0.088 | 0.748 |
3c | 0.453 | 0.080 | 0.711 |
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Stoean, B.; Rugina, D.; Focsan, M.; Craciun, A.-M.; Nistor, M.; Lovasz, T.; Turza, A.; Porumb, I.-D.; Gál, E.; Cristea, C.; et al. Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents. Int. J. Mol. Sci. 2021, 22, 2985. https://doi.org/10.3390/ijms22062985
Stoean B, Rugina D, Focsan M, Craciun A-M, Nistor M, Lovasz T, Turza A, Porumb I-D, Gál E, Cristea C, et al. Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents. International Journal of Molecular Sciences. 2021; 22(6):2985. https://doi.org/10.3390/ijms22062985
Chicago/Turabian StyleStoean, Bianca, Dumitrita Rugina, Monica Focsan, Ana-Maria Craciun, Mǎdǎlina Nistor, Tamas Lovasz, Alexandru Turza, Ioan-Dan Porumb, Emese Gál, Castelia Cristea, and et al. 2021. "Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents" International Journal of Molecular Sciences 22, no. 6: 2985. https://doi.org/10.3390/ijms22062985
APA StyleStoean, B., Rugina, D., Focsan, M., Craciun, A. -M., Nistor, M., Lovasz, T., Turza, A., Porumb, I. -D., Gál, E., Cristea, C., Silaghi-Dumitrescu, L., Astilean, S., & Gaina, L. I. (2021). Novel (Phenothiazinyl)Vinyl-Pyridinium Dyes and Their Potential Applications as Cellular Staining Agents. International Journal of Molecular Sciences, 22(6), 2985. https://doi.org/10.3390/ijms22062985