CN116621759A - 一种双重检测过氧亚硝基和二氧化硫的荧光探针及其应用 - Google Patents
一种双重检测过氧亚硝基和二氧化硫的荧光探针及其应用 Download PDFInfo
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- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
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- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/56—Ring systems containing three or more rings
- C07D209/58—[b]- or [c]-condensed
- C07D209/60—Naphtho [b] pyrroles; Hydrogenated naphtho [b] pyrroles
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Abstract
本发明提供了一种双重检测过氧亚硝基和二氧化硫的荧光探针及其应用,探针以6‑二甲胺基‑2‑萘甲醛为主体,在此结构上分别接上两个类似的吲哚盐衍生物,制备出具有红色发射的探针NAI和探针NABI。光谱结果证明了两种荧光探针对二氧化硫、ONOO‑、以及亚硫酸钠均有明显的响应效果。本发明提供的荧光探针能够同时区分检测过氧亚硝基阴离子、二氧化硫,检测限低,响应时间短,并对细胞内粘度、RNA均有很好的响应效果,能有效检测细胞凋亡过程。
Description
技术领域
本发明属于有机分子荧光探针技术领域,具体涉及一种双重检测过氧亚硝基和二氧化硫的荧光探针及其应用。
背景技术
细胞凋亡是细胞的一种自杀模式,利用这种模式去处理受到损伤或被判断为对生物体可能产生威胁的细胞。研究发现,生物体内每天要产生大量的凋亡细胞以维持系统平衡。细胞凋亡时,会出现细胞收缩、染色质凝结等特征。细胞凋亡异常容易导致多种疾病,如神经退行性疾病;程度严重的甚至会变成癌症,如艾滋病。因此,迫切需要开发监测细胞凋亡的方法。其中,荧光成像经常因原位、实时、高灵敏度等优势被用来监控细胞凋亡。
过氧亚硝酸盐ONOO-是一种活性氧,其半衰期非常短,不到十毫秒。ONOO-在生物体内的信号转导和杀菌反应中有重要作用。但是一旦ONOO-过量,会因其本身的氧化性、亲核性、硝化性去破坏细胞。此外,ONOO-可以诱发许多常见疾病。因此,检测ONOO-成为研究热点。荧光探针因其灵敏度高、操作简便等特点,成为检测ONOO-的有力工具之一。目前,设计检测ONOO-的荧光探针的思路有很多。比如开发以苯硼酸酯基或肼基为反应位点的荧光探针。
二氧化硫是一种常见的活性氧,当二氧化硫溶解在水中,形成的盐会被氧化成硫酸,而硫酸就是酸雨的主要组成部分,其对河流会造成严重污染。在生物系统中,内源性二氧化硫是可以在一些含硫氨基酸的氧化过程中产生,其在维持生物系统中氧化还原状态发挥着重要作用。研究发现,吸入过量的二氧化硫会诱发一些呼吸道疾病。因此,开发能够检测二氧化硫的方法十分重要。常见的检测二氧化硫的方法有很多,如分光光度法、高效液相色谱法、电化学法等,但一般操作复杂。荧光探针相对来说操作较为简便。
根据研究发现,二氧化硫、ONOO-和细胞凋亡这三者存在着微妙的联系。ONOO-能够与蛋白质反应,严重时会导致细胞凋亡;二氧化硫能够抑制血管平滑肌细胞增殖,促进细胞凋亡。然而,能够同时观察这三者之间关系的荧光探针却少有报道。
发明内容
本发明针对现有荧光探针的不足设计、合成了两种能够对二氧化硫、ONOO-区分产生荧光信号变化的荧光探针,分别是探针NAI和探针NABI。光谱结果证实了探针NAI和探针NABI对二氧化硫和ONOO-的响应能力,并且能够通过发射波长进行有效区分。成像实验研究发现,探针NABI对细胞凋亡有监控能力。本发明公开的一种区分检测过氧亚硝基阴离子和二氧化硫的荧光探针具有以下NAI和NABI结构:
本发明荧光探针NAI和NABI的制备方法如下:
化合物1的合成:在室温下,向加有2,3,3-三甲基-3H-吲哚的乙醇溶液中加入碘甲烷。将混合溶液加热到80℃并反应12小时,然后冷却到室温。将固体过滤,用乙醇洗涤,然后干燥,得到白色产物。
化合物2的合成:在室温下,向含有1,1,2-三甲基-1H-苯并吲哚的乙醇溶液中加入碘甲烷。将混合溶液加热到80℃并反应12小时,然后冷却到室温。将粗产物过滤,用乙醇洗涤,然后干燥,得到白色固体。
探针NAI的合成:化合物1和6-二甲氨基-2-萘醛加入到含有2mL乙醇的密封管中。在密封管中搅拌加入哌啶。系统加热到80℃反应10小时,然后冷却到室温。过滤得到粗产物,经乙醇重结晶后得到棕色固体。
探针NABI的合成:化合物2和6-二甲氨基-2-萘醛加入到含有2mL乙醇的密封管中。在密封管中搅拌加入哌啶。系统加热到80℃反应10小时,然后冷却到室温。过滤得到粗产物,经乙醇重结晶后得到棕色固体。
本发明公开了上述检测过氧亚硝基阴离子、二氧化硫的荧光探针在细胞成像可视化的应用。具体的,所述细胞成像为细胞凋亡成像、细胞活性成像,
进一步优选的,所述细胞成像为对干细胞、内质网或者癌症细胞成像。
进一步优选的,本发明所述荧光探针能同时检测细胞内ONOO-、二氧化硫与亚硫酸钠。
进一步优选的,上述制备的荧光探针NAI和NABI能够对细胞内粘度、RNA积极响应。
进一步优选的,上述荧光探针能与细胞内RNA结合。
进一步优选的,上述荧光探针在制备神经退行性疾病、癌症检测试剂的应用
本发明荧光探针可以用于细胞成像的主要原因是该荧光探针能检测细胞内ONOO-以及二氧化硫浓度变化,并且检测灵敏。同时本发明的荧光探针对细胞内粘度、RNA均有积极响应。
本发明的有益效果:
吲哚盐化合物NAI和NABI可同时区分检测过氧亚硝基阴离子和二氧化硫,并且能够通过发射波长进行有效区分;响应时间短,在1分钟内即可反应完全,响应能力良好;探针NAI和NABI对粘度、RNA均有很好的响应效果;探针NAI和NABI能很好的检测细胞凋亡过程。
附图说明
图1为实施例1制备的化合物NAI的1H NMR谱图;
图2为实施例1制备的化合物NABI的1H NMR谱图;
图3为探针NABI和探针NAI对亚硫酸钠浓度滴定荧光光谱测试图;
10μM NABI在400nm(A)和560nm(B)激发下,关于亚硫酸钠的荧光光谱图;10μMNABI在400nm(C,D)和560nm(E,F)激发下,关于200μM亚硫酸钠的动力学荧光光谱图。
图4为探针NABI和探针NAI对亚硫酸钠动力学光谱测试图;
10μM NAI在365nm(A)和560nm(B)激发下,关于亚硫酸钠的荧光光谱图;10μM NAI在365nm(C,D)和560nm(E,F)激发下,关于100μM亚硫酸钠的动力学荧光光谱图。
图5为ONOO-浓度滴定荧光光谱测试图,
10μM NABI在400nm(A)和560nm(B)激发下,关于ONOO-的荧光光谱图;10μM NABI在400nm(C,D)和560nm(E,F)激发下,关于600μM的ONOO-的动力学荧光光谱图。
图6为探针NABI和探针NAI对ONOO-动力学光谱测试图;
10μM NAI在400nm(A)和560nm(B)激发下,关于ONOO-的荧光光谱图;10μM NAI在400nm(C,D)和560nm(E,F)激发下,关于600μM的ONOO-的动力学荧光光谱图。
图7为探针NABI和探针NAI对粘度荧光光谱测试图;
(A)10μM NABI在560nm激发下,在甲醇和甘油不同比例组成的混合溶液中的荧光光谱;(B)10μM NABI关于RNA滴定浓度的吸收光谱图;(C)10μM NABI在560nm激发下,关于RNA滴定浓度的荧光光谱图。
图8为4.3.7RNA浓度滴定紫外、荧光光谱测试图;
(A)10μM NAI在560nm激发下,在甲醇和甘油不同比例组成的混合溶液中的荧光光谱;(B)10μM NAI关于RNA滴定浓度的吸收光谱图;(C)10μM NAI在560nm激发下,关于RNA滴定浓度的荧光光谱图。
图9为NABI选择性荧光光谱测试图;
10μM NABI在400nm(A)和560nm(B)激发下,在未加入干扰物和加入600μM干扰物的荧光光谱图。
图10为NAI选择性荧光光谱测试图;
10μM NAI在365nm(A)和400nm(B)和560nm(C)激发下,在未加入干扰物和加入600μM干扰物的荧光光谱图。
图11为固定细胞和用DNase处理或RNase处理的固定细胞用NABI孵育30分钟图像;
图12为探针NABI在固定细胞和用500μM的二氧化硫或500μM的ONOO-处理的固定细胞荧光图。
具体实施方式
为了使本发明的目的、技术方案更加清楚明白,以下结合实施例,对本发明做进一步的说明,但是本发明的保护范围并不限于这些实施例,实施例仅用于解释本发明。本领域技术人员应该理解的是,凡是不背离本发明构思的改变或等同替代均包括在本发明的保护范围之内。下面结合实施例对本发明进一步描述,但本发明并不为实施例所限制。
本发明所使用药品均属外购:6-二甲胺基-2-萘甲醛和1,1,2-三甲基-1H-苯并吲哚均购买于郑州阿尔法化工有限公司;2,3,3-三甲基-3H-吲哚购买于安徽泽升科技有限公司;碘甲烷购买于北京百灵威科技有限公司。生物成像所需试剂:DNA和RNA购自西格玛奥德里奇(上海)贸易有限公司(项目编号:DNA为D1501,RNA为R6625,它们是从小牛胸腺和酵母中分离出来的天然DNA和RNA)。
有机合成仪器:磁力搅拌器、油浴锅和真空泵(R5614Y-1)采购于巩义市予华仪器有限公司;旋转蒸发仪(N-1100)购买于东京理化公司;电子天平(LE204E/02)购买于梅特勒上海有限公司;生物成像仪器:Nikon A1MP共聚焦显微镜购买于Nikon公司;二氧化碳培养箱购买于松下电器有限公司。
实施例1:
化合物1的合成:在室温下,向加有2,3,3-三甲基-3H-吲哚(1.0g,6.28mmol)的乙醇溶液(3mL)中加入碘甲烷(1.0g,7.05mmol)。将混合溶液加热到80℃并反应12小时,然后冷却到室温。将固体过滤,用乙醇洗涤,然后在60℃下干燥,得到白色产物。(1.63g,产率86%)。
化合物2的合成:在室温下,向含有1,1,2-三甲基-1H-苯并吲哚(1.0g,4.78mmol)的乙醇溶液(3mL)中加入碘甲烷(0.81g,5.71mmol)。将混合溶液加热到80℃并反应12小时,然后冷却到室温。将粗产物过滤,用乙醇洗涤,然后在60℃下干燥,得到白色固体。(1.43g,产率85%)。
探针NAI的合成:化合物1(151mg,0.5mmol)和6-二甲氨基-2-萘醛(100mg,0.5mmol)加入到含有2mL乙醇的密封管中。在密封管中搅拌加入哌啶(2.1mg,25μmol)。系统加热到80℃反应10小时,然后冷却到室温。过滤得到粗产物,经乙醇重结晶后得到棕色固体(207.4mg,86%)。1H NMR(600MHz,DMSO-d6)δ8.58–8.45(m,2H),8.18(dd,J=8.8,1.1Hz,1H),7.85(t,J=9.0Hz,3H),7.77(d,J=8.8Hz,1H),7.65–7.53(m,3H),7.30(dd,J=9.1,2.4Hz,1H),7.03(d,J=2.2Hz,1H),4.11(s,3H),3.13(s,6H),1.81(s,6H).核磁氢谱见附图1。
探针NABI的合成:化合物2(176mg,0.5mmol)和6-二甲氨基-2-萘醛(100mg,0.5mmol)加入到含有2mL乙醇的密封管中。在密封管中搅拌加入哌啶(2.1mg,25μmol)。系统加热到80℃反应10小时,然后冷却到室温。过滤得到粗产物,经乙醇重结晶后得到棕色固体(233.6mg,84%)。1H NMR(600MHz,DMSO-d6)δ8.65–8.50(m,2H),8.43(d,J=8.1Hz,1H),8.28(d,J=8.6Hz,1H),8.21(s,2H),8.08(d,J=8.6Hz,1H),7.87(d,J=8.8Hz,1H),7.79(d,J=8.6Hz,2H),7.74–7.68(m,1H),7.63(d,J=16.1Hz,1H),7.31(d,J=8.3Hz,1H),7.04(s,1H),4.24(s,3H),3.11(d,J=31.1Hz,6H),2.04(s,6H).核磁氢谱见附图2。
实施例2:探针NAI和NABI光谱测试
探针NAI母液的配制:称取2.4mg的探针溶于1mL的DMSO中配成5mM的母液。
探针NABI母液的配制:称取2.6mg的探针溶于1mL的DMSO中配成5mM的母液。
亚硫酸钠的制备:称取12.6mg的亚硫酸钠溶于10mL的去离子水中配成10mM的溶液。
关于亚硫酸钠的荧光光谱测试方法:先配一个空白样,取4μL浓度为5mM的探针溶液加入到2mL PBS缓冲液和乙醇的混合溶液中(体积比为1比1),此时探针的工作浓度为10μM,接着加入一定浓度的亚硫酸钠溶液进行荧光的光谱测试。(探针NAI荧光光谱测试中激发波长为365nm和560nm,探针NABI荧光光谱测试中激发波长为400nm和560nm)
ONOO-的制备:在冰浴条件下,向剧烈搅拌的含有NaNO2(0.6M,10mL)和过氧化氢(0.7M,10mL)的混合溶液中加入HCl(0.6M,10mL),然后迅速加入氢氧化钠(1.5M,20mL)。最后,加入极少量的二氧化锰并过滤。注意,上述溶液均由去离子水制备,且制备好的ONOO-须在-20℃条件下避光保存。ONOO-浓度标定:先用2mL浓度为0.1M的氢氧化钠扫一遍基线,再往V mL的浓度为0.1M氢氧化钠里加X微升制备好的ONOO-溶液并进行紫外测定,用以下计算,其中A302 nm是ONOO-在302nm处的吸光度,V是溶液总体积,X是ONOO-溶液的体积。
关于ONOO-的吸收、荧光光谱测试方法:先配一个空白样,取4μL浓度为5mM的探针溶液加入到2mL PBS缓冲液和乙醇的混合溶液中(体积比为1比1),此时探针的工作浓度为10μM,接着加入一定浓度的ONOO-溶液进行紫外或荧光的光谱测试。(探针NAI和探针NABI荧光光谱测试中激发波长为400nm和560nm)
关于亚硫酸氢钠的吸收、荧光光谱测试方法:先配一个空白样,取4μL浓度为5mM的探针溶液加入到2mL PBS缓冲液和乙醇的混合溶液中(体积比为1比1),此时探针的工作浓度为10μM,接着加入一定浓度的亚硫酸氢钠溶液进行紫外或荧光的光谱测试。(探针NAI荧光光谱测试中激发波长为365nm和560nm,探针NABI荧光光谱测试中激发波长为400nm和560nm)
关于粘度的荧光光谱测试方法:配制好甘油和甲醇以不同体积占比组成的混合溶液,分别取2μL浓度为5mM的探针、探针溶液加入到2mL上述提及的不同比例的混合溶液中,即可进行荧光光谱测试。(荧光光谱测试中激发波长均为560nm)
关于RNA的荧光光谱测试方法:分别取2μL浓度为5mM的探针和探针溶液加入到2mL的PBS缓冲液中,接着分别加入一定量的RNA溶液进行荧光光谱测试。RNA摩尔浓度测定:最初配制浓度为1mg/mL的RNA溶液,为了量化它们的摩尔浓度,将部分原液稀释。RNA溶液稀释250倍。然后测量稀释后的RNA溶液的吸收光谱。(荧光光谱测试中激发波长均为560nm)
关于选择性荧光光谱测试方法:在这里选择半胱氨酸、谷胱甘肽、甘氨酸、丝氨酸、脯氨酸、丙氨酸、谷氨酸、苏氨酸、组氨酸、缬氨酸、天冬酰胺、苯丙氨酸、精氨酸、葡萄糖、蔗糖、氯化钠、氯化钾、硝酸钾、过氧化氢、VC(工作浓度均为600μM)为待测干扰物质,称取一定量的上述分析物质,并用去离子水溶解。取4μL浓度为5mM的探针溶液加入到2mL PBS缓冲液和乙醇的混合溶液中(体积比为1比1),接着分别加入上述干扰物质进行荧光的光谱测试。(探针NAI荧光光谱测试中激发波长为365nm、400nm、560nm,探针NABI荧光光谱测试中激发波长为400nm、560nm)
(1)亚硫酸钠浓度滴定荧光光谱测试:
为了验证探针NABI和探针NAI对亚硫酸钠的响应能力,对这两种探针进行了关于亚硫酸钠浓度滴定的荧光光谱测试。首先探针NABI的激发波长在400nm,探针NAI的激发波长在365nm。如附图3中A和附图4中A所示,未加入亚硫酸钠时,探针NABI和探针NAI基本上没有荧光;滴加亚硫酸钠之后,逐渐检测到探针NABI和探针NAI的荧光信号。当分别加入2个或1个当量的亚硫酸钠时,探针NABI或探针NAI的荧光强度不再变化,说明此时达到饱和浓度。探针NABI荧光强度从最初的38增加到5500,变化了近145倍;探针NAI荧光强度从最初的38增加到1055,变化了近28倍。探针NABI和探针NAI荧光信号增强说明这两种探针均在不断反应,探针NABI生成了一种发射波长大概在480nm左右的新的物质,探针NAI则是在437nm。为了反向验证,在560nm激发下,重复进行了上述实验。如附图3中B和附图4中B所示,探针NABI和探针NAI在未加入亚硫酸钠时,探针NABI本身的发射波长大约在735nm左右,探针NAI在715nm左右;当逐渐加入亚硫酸钠后,探针NABI和探针NAI在该发射峰的荧光强度不断降低,说明两种探针均在不断与亚硫酸钠反应。当分别加入荧光探针各自的亚硫酸钠饱和浓度,探针NABI和探针NAI在发射峰的荧光信号检测不到,说明此时反应结束。
(2)亚硫酸钠动力学光谱测试:
为了了解探针NABI和探针NAI对亚硫酸钠的响应速度,进行了其关于亚硫酸钠的动力学测试。如附图3中C、D,附图4中C、D所示,分别在400nm和365nm激发下,探针NABI和探针NAI本身荧光度十分微弱;分别加入亚硫酸钠饱和浓度后,荧光强度均快速变化,并且很快保持不变。同样的,从附图3中E、F、附图4中E、F看到,在560nm激发下,探针NABI和探针NAI本身荧光较强;加入亚硫酸钠饱和浓度后,荧光强度均立马降低。探针NABI对亚硫酸钠的响应时间大概在4分钟,探针NAI则是1分钟。
(3)ONOO-浓度滴定荧光光谱测试:
为了确定探针NABI和探针NAI对ONOO-是否可以响应,进行了其关于ONOO-的浓度滴定荧光光谱测定。如附图5中A、附图6中A所示,在400nm激发下,探针NABI和探针NAI本身的荧光信号十分微弱,几乎检测不到;加入ONOO-之后,探针NABI和探针NAI的荧光信号均发生了明显的增强;且加入60个当量的ONOO-之后,荧光强度均不再变化。探针NABI荧光强度从26增强到1209,变化了近46倍;探针NAI荧光强度从31增强到1598,变化了近50倍。整体来看,两种探针均对ONOO-响应良好。为了反向验证ONOO-反应情况,在560nm激发下,重复上述实验内容。如图附图5中B、附图6中B所示,未加ONOO-时,探针NABI和探针NAI的荧光信号很强;加入ONOO-后其荧光信号均不断减弱,并在加入ONOO-饱和浓度后荧光信号均非常微弱,说明反应结束。
(4)ONOO-动力学光谱测试:
为了进一步了解探针NABI和探针NAI对ONOO-的响应灵敏度,进行了其关于ONOO-的动力学光谱测试。如图附图5中C、D,附图6中C、D所示,在400nm激发下,未加ONOO-时,探针NABI和探针NAI本身荧光很弱;加入ONOO-饱和浓度之后,探针荧光均快速增强并保持不变。整个过程探针NABI只需0.8分钟,探针NAI只需1分钟。同样的,如附图5中E、F,附图6中E、F所示,在560nm激发下,探针NABI和探针NAI本身荧光很强;加入ONOO-之后,荧光迅速减弱并很快变为最低。整体来看,探针NABI和探针NAI对ONOO-的响应时间均非常短。
(5)粘度荧光光谱测试:
为了研究探针NABI和探针NAI对粘度的响应情况,进行了其关于粘度的光谱测试。如附图7中A、附图8中A所示,在560nm激发下,探针NABI和探针NAI本身的荧光强度较低;当测试体系粘度增加时,两种探针的荧光均明显增强。当体系粘度从0.6cp逐渐增加到9453cp时,探针NABI的荧光强度从最初的222增加到3064,变化了近14倍;探针NAI的荧光强度从最初的152增加到3062,变化了近20倍;由此判断两种探针均对粘度响应良好。
(6)RNA浓度滴定紫外、荧光光谱测试
为了了解探针NABI和探针NAI关于RNA的光学性能,首先进行了其关于RNA的紫外浓度滴定光谱测试。如附图7中B、附图8中B所示,探针NABI未加入RNA和加入RNA时的最大吸收峰没有太大区别,波长范围在450nm-600nm,大概位于530nm左右;探针NAI则是在该波长范围内的525nm左右。接着,又进行了探针NABI和探针NAI关于RNA浓度滴定荧光光谱测试。如附图7中C、附图8中C所示,在560nm激发下,探针NABI在加入RNA后,荧光信号在720nm左右明显增强,当加入20个当量的RNA后,荧光强度达到最大值;探针NAI在加入RNA后,荧光信号在710nm左右明显增强,当加入50个当量的RNA后,荧光强度达到最大值;上述结果表明了两种探针对RNA均有一定的亲和力。
(7)选择性荧光光谱测试:
由于设计出的探针需要在细胞内进行孵育以进行细胞成像实验,而细胞内存在着多种生物小分子,如果探针与其进行响应,就会干扰成像实验结果甚至得出错误的实验结论,这就需要设计出的探针具备良好的选择性,这样成像实验结果才更加有说服力。如附图9、附图10所示,在400nm、560nm激发下,探针NABI对这些干扰物均不响应,荧光强度与空白对照组相比几乎没有变化;在365nm、400nm、560nm激发下,探针NAI对这些干扰物均不响应,荧光强度与空白对照组相比几乎没有变化;这很好的为下面的生物成像实验奠定了基础。
实施例3:细胞成像实验
(1)固定细胞成像实验
为了了解探针NABI在活细胞和固定细胞中的染色及分布情况,进行了相关的细胞成像实验。如附图11所示,活细胞中,探针NABI在红通道发射强烈,主要分布在细胞质中,荧光信号呈丝状表达。固定细胞中,探针在红通道中的荧光信号增强,并分布在细胞质和核仁中。固定细胞用DNase处理后,探针在红通道的荧光信号没有变化,说明探针未与DNA结合。固定细胞用RNase处理后,红色荧光信号几乎消失,说明探针与RNA结合。
(2)二氧化硫及ONOO-细胞成像实验
为了了解探针NABI在固定细胞中用二氧化硫及ONOO-处理后的染色及分布情况,进行了相关的细胞成像实验。如附图12所示,在细胞未经处理时,探针NABI在近红外通道中产生明显的荧光信号,信号位于细胞核中;当细胞用二氧化硫处理后,探针在近红外通道中的荧光信号明显减弱,并在蓝色通道中出现了明显的荧光信号;当细胞用ONOO-处理后,探针在近红外通道中的荧光信号减弱,并在绿色通道中出现了明显的荧光信号。结果表明,NABI探针在固定细胞中加入二氧化硫或ONOO-之后均有良好的响应。
Claims (7)
1.一种双重检测过氧亚硝基和二氧化硫的荧光探针,所述荧光探针为吲哚盐化合物,其特征在于,所述荧光探针代号分别为NAI,NABI,其中荧光探针NAI,NABI具有如下结构式:
2.根据权利要求1所述的双重检测过氧亚硝基和二氧化硫的荧光探针在细胞成像可视化的应用。
3.根据权利要求2所述的细胞成像可视化的应用,其特征在于,所述细胞成像为细胞凋亡成像、细胞活性成像。
根据权利要求2所述的细胞成像可视化的应用,其特征在于,所述细胞成像为对干细胞、内质网或者癌症细胞成像。
4.根据权利要求2所述的细胞成像可视化的应用,其特征在于,所述荧光探针能同时检测细胞内ONOO-、二氧化硫与亚硫酸钠。
5.根据权利要求2所述的细胞成像可视化的应用,其特征在于,所述荧光探针对细胞内粘度、RNA积极响应。
6.根据权利要求5所述的细胞成像可视化的应用,其特征在于,所述荧光探针能与细胞内RNA结合。
7.根据权利要求1所述的双重检测过氧亚硝基和二氧化硫的荧光探针在制备神经退行性疾病、癌症检测试剂的应用。
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