二氟化硼β-二酮类近红外荧光染料的设计、合成及细胞成像

doi:10.6023/cjoc202404032

摘要/Abstract

近红外(NIR)发光材料因具有生物组织穿透力强和光散射弱的独特优势在生物成像领域具有广泛应用的潜力. 通过增强分子内电荷转移策略设计合成了具有供体-受体(D-A)结构的二氟化硼β-二酮配合物(TPA-BF₂-CN). TPA-BF₂-CN具有溶剂依赖发光特性, 在稀溶液和固态下均具有较好的发光效率, 分别为11.1%和5.9%, 荧光发射波长最大峰值达760 nm, 处于NIR范围; 具有力致荧光变色性质, 研磨前后发射波长可红移107 nm; 还表现出双发射荧光现象, 两个发射峰分别位于400 nm的蓝光区和760 nm的NIR区, 其中NIR区的起止发射波长范围为690~850 nm. 相比于单通道荧光分子更具有避免环境因素干扰的优势. 细胞成像研究表明, TPA-BF₂-CN对HeLa细胞相容性良好, 并且在整个HeLa细胞质区域中可观察到近红外发射信号. 因此, TPA-BF₂-CN可作为一种活细胞成像荧光剂.

关键词: 二氟化硼β-二酮, 近红外, 细胞成像, 力致变色, 双态发射

Near-infrared (NIR) luminescent materials have great potential in the field of bioimaging due to their unique advantages of strong tissue penetration and weak light scattering in biological tissues. In this study, a donor-acceptor (D-A) structured boron difluoride β-diketonate complex (TPA-BF₂-CN) was designed and synthesized through the strategy of enhancing intramolecular charge transfer. This complex exhibits solvent-dependent luminescent properties. It demonstrates good luminescence efficiency in dilute solution and solid state with efficiencies of 11.1% and 5.9%, respectively. The maximum emission wavelength peaks at 760 nm, falling within the NIR range. The complex also possesses mechanochromic properties with a redshift of emission wavelength by 107 nm after grinding. TPA-BF₂-CN exhibits dual emission fluorescence phenomenon with emission peaks located in the blue light region at 400 nm and the NIR region at 760 nm with an emission wavelength range of 690~850 nm in the NIR region. Compared to single-channel fluorescent molecules, it has the advantage of avoiding interference from environmental factors. Cell imaging studies demonstrate good compatibility of TPA-BF₂-CN with HeLa cells, and NIR emission signals can be observed throughout the cytoplasmic region of HeLa cells. Therefore, TPA-BF₂-CN can serve as a fluorescent probe for live cell imaging.

Key words: difluoroboron β-diketonate, near-infrared, cell imaging, mechanochromic, dual-state emission

引用此文

齐云鹏, 刘伟, 王俊龙, 杜涛, 矫淞霖. 二氟化硼β-二酮类近红外荧光染料的设计、合成及细胞成像[J]. 有机化学, 2024, 44(11): 3357-3364.

Yunpeng Qi, Wei Liu, Junlong Wang, Tao Du, Songlin Jiao. Design and Synthesis of Near-Infrared Fluorescent Dyes of Difluoroboron β-Diketonate and Cell Imaging[J]. Chinese Journal of Organic Chemistry, 2024, 44(11): 3357-3364.

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图/表 10

图式1. TPA-BF2-CN的合成路线

Scheme 1. Synthetic route to the TPA-BF2-CN

In solution						Pristine powder				Ground powder
Solvent	λ_abs^a/nm	ε^b/(L•mol^－1•cm^－1)	λ_em^c/nm	Stokes shift	Φ_f^d/%	τ^e/ns	λ_em^f/nm	λ_em^g/nm	Φ_f^d	Φ_f^d/%	λ_em^f/nm	λ_em^g/nm
Hexane	471	8660	539	2678	10.0
Toluene	484	44310	598	3938	11.1
THF	485	32590	588	3611	5.1	3.09	627	396/760	5.9	0.2	695	397/698
DCM	494	58150	620	4113	2.3
DMSO	495	35640	603	3618	2.2

表1. TPA-BF2-CN在各种状态下的光物理数据

Table 1. Photophysical data of TPA-BF2-CN in various states

图1. TPA-BF2-CN在不同有机溶剂中的归一化紫外吸收光谱

Figure 1. Normalized UV-vis spectra of TPA-BF2-CN in various solvents

图2. TPA-BF2-CN在不同有机溶剂中的归一化荧光发射光谱(A)和TPA-BF2-CN在不同溶剂中的照片(B)

Figure 2. Normalized fluorescence spectra of TPA-BF2-CN in various solvents (A), and photographs of TPA-BF2-CN in various solvents (B)

图3. TPA-BF2-CN的HOMO和LUMO能级的分子轨道分布图

Figure 3. Molecular orbital amplitude plots of HOMO and LUMO levels for TPA-BF2-CN

图4. TPA-BF2-CN在THF/水混合体系中的荧光发射光谱(A)和TPA-BF2-CN在THF/水混合体系中于365 nm紫外光照射下的图片(B)

Figure 4. Fluorescence emission spectra of TPA-BF2-CN in the THF/water mixtures with different fractions of water (A), and photographs in THF/water mixtures of TPA-BF2-CN under 365 nm UV illumination (B)

图5. TPA-BF2-CN的甘油溶液在不同温度下的荧光发射光谱

Figure 5. Fluorescence emission spectra of TPA-BF2-CN in glycerol solution at different temperatures Insert: trends of fluorescence intensity changes of TPA-BF2-CN in glycerol solution at different temperatures.

图6. 固态的TPA-BF2-CN在490 nm激发光下的荧光发射光谱(A)、固态的TPA-BF2-CN在320 nm激发光下的荧光发射光谱(B)、TPA-BF2-CN在不同固态下的XRD谱图(C)、TPA-BF2-CN在不同固态下于紫外灯下的照片(D)

Figure 6. Fluorescence emission spectrum of solid TPA-BF2-CN under 490 nm excitation (A), fluorescence emission spectrum of solid TPA-BF2-CN under 320 nm excitation (B), XRD spectrum of TPA-BF2-CN in different solid states (C), and photographs of TPA-BF2-CN in different solid states under UV light (D)

图7. 采用MTT试验评估TPA-BF2-CN对HeLa细胞的毒性

Figure 7. Toxicity of TPA-BF2-CN to HeLa cells using the MTT assay

图8. TPA-BF2-CN孵育后的HeLa细胞的CLSM图像

Figure 8. CLSM images of HeLa cells incubated with TPA-BF2- CN (A) Images in blue channel (λex=405 nm, λem=410~466 nm); (B) Images in red channel (λex=405 nm, λem=626~672 nm); (C) Images in green channel (λex=488 nm, λem=597~624 nm); (D) Images in bright field channel; (E) Overlay of A, B, C, D, and F; (F) Images in NIR channel (λex=488 nm, λem=626~663 nm).

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