线粒体荧光探针最新研究进展

细胞器的研究一直是认识细胞结构和功能的重要手段。线粒体是细胞的能量工厂,参与众多的新陈代谢过程,许多病理学过程均与它相关,是一种重要的细胞器,一直以来都是研究的热点。定位于线粒体的荧光探针主要分为3大类,本文重点介绍并讨论近10年基于带有正电荷的荧光团或者在荧光团中引入三苯基膦等定位基团实现对线粒体染色荧光探针的研究进展。     

识别细胞内金属离子的有机荧光探针研究进展

金属离子广泛存在于组织细胞和体液中,在人体的生理和病理中发挥着十分重要的作用。金属离子的浓度必须精确控制在一定的范围内,某些离子浓度的微弱变化都将引发人体的疾病。由于细胞是生命活动的基本单位,因此研究各种金属离子对生物体的影响,通过对活体细胞内金属离子的荧光显微成像进行细胞内离子的可视化定量和定性分析引起了人们的极大关注。就近年来人们对识别细胞内金属离子的有机荧光探针与细胞成像研究领域的进展进行了评述。     

β-半乳糖苷酶荧光探针的研究进展

β-半乳糖苷酶属于糖苷水解酶,在食品工业、基因工程、酶工程、蛋白质工程、生物医学等领域发挥着重要作用。β-半乳糖苷酶活性及含量的异常与癌症的发生发展密切相关,因此该酶检测对早期癌症诊断有着重要意义。传统酶活性检测方法具有一定的局限性,无法实现对β-半乳糖苷酶的原位无损检测。荧光探针因灵敏度高、选择性好、可用显微镜直接观察、分辨率高、响应时间短、操作方便、成本低等优点深受研究者青睐。在简述荧光探针设计原理的基础上,对近年来β-半乳糖苷酶荧光探针的设计合成研究进展进行了综述,阐述了β-半乳糖苷酶荧光探针在生物成像、早期癌症诊断等应用进展,为β-半乳糖苷酶高效检测技术的研究提供了帮助。     

荧光探针在游离肼检测中的研究进展

肼是20世纪40年代火箭的经典推进剂,同时被广泛用作塑料发泡剂.此外,肼由于其还原性和碱性,还经常用于合成油漆、药物和杀虫剂.然而,肼对人体具有相当大的毒性作用,包括对眼睛和皮肤的刺激、对肝脏、肾脏和中枢神经系统的损害.因此对肼进行准确测定意义重大.荧光测定法由于其无创性和良好的灵敏度,更适合生物应用.介绍了肼探针的最...     

检测苯硫酚荧光探针的研究进展

苯硫酚(PhSH)是一类高活性和毒性的芳香族硫醇,在化学工业中广泛应用于制备农药、聚合物和药物。就近年来用于PhSH检测和成像的荧光探针的研究进展进行综述。这些探针根据识别部分进行了分类,并根据其结构和传感性能进行了详细介绍。此外,还讨论了未来的研究前景。     

靶向细胞器的过氧亚硝酸盐荧光探针研究进展

过氧亚硝酸盐（ONOO^-）作为氧代谢的重要产物,在免疫应答、细胞信号转导等许多生理活动中起着重要作用。然而,ONOO^-的水平异常会损伤细胞生物分子,与许多疾病密切相关。因此,迫切需要监测活细胞中ONOO^-的含量变化,而ONOO^-的功能很大程度上依赖于亚细胞区,所以开发亚细胞的ONOO^-水平变化的监测手段更有研究价值。为此,多种具有靶向细胞器的ONOO^-荧光探针被设计、开发和应用。重点介绍了近6年来具有线粒体、溶酶体和内质网靶向能力的ONOO^-荧光探针的代表性例子。综述了这些荧光探针的细胞器靶向策略、结构、荧光行为和生物学应用。     

反应激活型酶荧光探针的研究进展

酶在维持生物体内稳态与生命活动的正常运行方面发挥着举足轻重的作用。某些特定酶含量及活性的异常与人类重大疾病的发生与发展密切相关。因此,生物体内特定酶的实时原位检测及可视化成像具有重要的意义。化学荧光探针具有选择性好、灵敏度高及高时空分辨率可视化成像等优点,近年来研究者设计合成了大量的可用于生物体系内酶识别与可视化成像的荧光探针。目前识别酶的荧光探针主要有两类:(1)基于酶对荧光探针分子中酶抑制剂基团的识别引起探针荧光信号的变化;(2)基于酶对荧光探针特异性催化反应来实现识别前后荧光信号的激活,称为反应激活型酶荧光探针。对反应激活型酶荧光探针的设计策略及4种重大疾病相关的生物标志酶(单胺氧化酶、β-半乳糖苷酶、硝基还原酶、γ-谷氨酰转肽酶)的识别可视化荧光探针研究进展进行了综述,对未来酶识别荧光探针的研究方向进行了展望。     

荧光探针用于细胞内硒代半胱氨酸检测的研究进展

硒代半胱氨酸(Sec)在人体调控免疫反应、抑制癌症的发生,以及维持生理氧化还原平衡等方面发挥着十分重要的作用。采用荧光探针直接在分子水平上对硒代半胱氨酸进行体内实时无损检测是探索生理活动机制的最有效的方法。总结了不同类型的荧光探针分子的设计原理及其对应的传感检测机制和成像应用。对比了不同的分析检测方式以及效果,介绍了用于生物硒代半胱氨酸检测的特异性荧光探针的最新进展。     

荧光探针的研究进展与应用

由于其制备简单、灵敏度和选择性,荧光探针已成为广泛研究和工业领域的首选分析工具,有助于快速检测感兴趣的化学物质以及在细胞水平上研究重要的生理和病理过程。此外,开发的许多长波长荧光探针也被证明适用于体内生物医学应用,包括荧光引导的疾病诊断和治疗诊断学(例如,荧光前药)。用于检测离子、有机小分子和生物大分子(包括酶、DNA/RNA、脂质和碳水化合物)的传感试剂和材料的开发取得了令人印象深刻的进展,这些物质在生物和疾病相关事件中起着至关重要的作用。在这里我们主要介绍用于生物应用的荧光探针和功能材料的示例,为用于基础生物学研究和临床转化的强大荧光化学工具的未来发展提供新思路。     

反应型HClO/ClO-荧光探针的研究进展

次氯酸/次氯酸根是活性氧化物中重要的信号分子,在生物医药和环境安全等方面发挥着重要作用。反应型荧光探针基于出色的灵敏度、实时成像和生物兼容性好等特点,广泛应用于HClO/ClO^-的检测。本文评述了近5年反应型HClO/ClO^-荧光探针的研究进展,包括反应型HClO/ClO^-荧光探针的设计策略和识别机理,并从HClO/ClO^-引发的反应机理角度（碳碳双键的反应、硫族化合物的反应、醛肟基团的反应、腙/席夫碱的反应、酰肼/磺酰肼的反应、N,N-二甲基硫代氨基甲酸酯的反应和苯硼酸/硼酸酯的反应）概述了反应型HClO/ClO^-荧光探针的特点和实际应用,指出反应型HClO/ClO^-荧光探针的发展方向是合成性能优异的识别基团,构建选择性好、水溶性好、低荧光背景、光化学性能稳定和生物毒性低的反应型近红外HClO/ClO^-荧光探针,实现对生物体内外HClO/ClO^-的可视化检测及机理探索。     

Cell‐Cycle Markers and Biosensors

Since the first schematic illustrations of dividing cells, we have come a long way in characterising eukaryotic cells and defining their cell‐cycle status thanks to a number of complementary approaches. Although most of these approaches rely on cell‐fixation procedures to identify molecular components in cell lysates, cultured cells or tissues, the development of GFP technology has enabled visualisation of virtually any fusion protein in cellulo and in vivo, and the exploitation of functional elements with well‐defined spatiotemporal characteristics has enabled the development of genetically encoded fluorescent markers of cell‐cycle phases, thus providing novel means of characterising the status of living cells in real time with high resolution. Together with technological advances in fluorescence chemistry and imaging approaches, the more recent development of fluorescent biosensors has provided direct means of probing cell‐cycle regulators and of studying their dynamics with high spatial and temporal resolution. Here we review classical approaches that rely on cell fixation to characterise the cell‐cycle status and its regulatory enzymes, and we describe the more recent development of cell‐cycle markers based on genetically encoded fusions of fluorescent proteins with characteristic cell‐cycle features, and of fluorescent biosensor technology to probe cell‐cycle regulators in living cells. Biosensors not only provide a means of characterising the behaviour of cell‐cycle regulators in their natural environment, they are also very useful for comparative studies of biological processes in healthy and pathological conditions, and can be further applied to diagnostic approaches to assess the status of a specific target, and to monitor response to therapeutic intervention.     

Modulation of Cellular Fate of Vinyl Triarylamines through Structural Fine Tuning: To Stay or Not To Stay in the Mitochondria?

Mitochondria are involved in many cellular pathways and dysfunctional mitochondria are linked to various diseases. Hence efforts have been made to design mitochondria-targeted fluorophores for monitoring the mitochondrial status. However, the factors that govern the mitochondria-targeted potential of dyes are not well-understood. In this context, we synthesized analogues of the TP-2Bzim probe belonging to the vinyltriphenylamine (TPA) class and already described for its capacity to bind nuclear DNA in fixed cells and mitochondria in live cells. These analogues ( TP-1Bzim, TPⁿ-2Bzim, TP¹⁺-2Bzim, TN-2Bzim ) differ in the cationic charge, the number of vinylbenzimidazolium branches and the nature of the triaryl core. Using microscopy, we demonstrated that the cationic derivatives accumulate in mitochondria but do not reach mtDNA. Under depolarisation of the mitochondrial membrane, TP-2Bzim and TP¹⁺-2Bzim translocate to the nucleus in direct correlation with their strong DNA affinity. This reversible phenomenon emphasizes that these probes can be used to monitor ΔΨ_m variations.     

Targeted Delivery of an Activatable Fluorescent Probe for the Detection of Furin Activity in Living Cells

Furin, a protein convertase, plays a crucial role in the progression of tumors. In this work, a new fluorescent probe consisting of a peptide, Arg‐Val‐Arg‐Arg (RVRR), and an aminoluciferin fluorophore was designed and prepared for the responsive and activatable detection of furin activity in vitro and in living cells. We demonstrated that this probe could be responsive toward furin with an “off–on” florescence signal and generated an approximately 3.58‐fold enhancement in the fluorescence intensity in vitro. Fluorescence imaging in MDA‐MB‐468 and LoVo cells showed that the probe could be cleaved by overexpressed furin with fluorescence turn‐on in MDA‐MB‐468 cells, and this probe was also found to be capable of discriminating between furin‐overexpressing and furin‐deficient cell lines. Our research indicates that this probe has great potential for the detection of furin activity in living cells.