CN113834802B - 镧系金属掺杂碳量子点、镧系金属掺杂碳量子点-核酸适配体偶联物探针的制备方法及应用 - Google Patents
镧系金属掺杂碳量子点、镧系金属掺杂碳量子点-核酸适配体偶联物探针的制备方法及应用 Download PDFInfo
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Abstract
本发明公开了一种镧系金属掺杂碳量子点的制备方法,包括如下步骤:将尿素和镧系金属盐Ln(NO3)3·nH2O,溶解于N,N‑二甲基甲酰胺中,室温下磁力搅拌;向上述溶液加入柠檬酸,溶解后转移至聚四氟乙烯内衬管内并放入不锈钢反应釜中;将拧紧的所述不锈钢反应釜放入马弗炉反应后自然冷却至室温;样品溶液转移至离心管内,离心,取上清液,得到沉淀物;将所述沉淀物中加入乙醇,超声溶解,离心,取沉淀;将所述沉淀用去离子水溶解,用透析袋透析;旋蒸浓缩,冻干得到黑色粉末,于干燥箱保存,得到CDs(Ln)。本发明还提供镧系金属掺杂碳量子点‑核酸适配体偶联物的制备方法及在荧光显微成像和质谱流式成像中的应用。
Description
技术领域
本发明涉及质谱成像金属标记探针技术领域,尤其涉及一种镧系金属掺杂碳量子点、镧系金属掺杂碳量子点-核酸适配体偶联物探针的制备方法及应用。
背景技术
质谱流式成像技术(Imaging Mass Cytometry,IMC)融合了免疫染色和质谱技术,能够高度精确地鉴别多种不同的同位素。IMC技术已经被用于组织切片或细胞爬片上超过三十个靶标分子的同时检测。然而,IMC技术也受限于其成像速度慢以及成像分辨率不高,激光烧蚀1mm2的区域大致需要80分钟。因此,只对感兴趣的区域进行IMC分析将大大提高IMC使用效率,若能通过荧光显微成像的快速扫描预先确定感兴趣区域,这将大大节省IMC盲扫时间。然而目前尚未开发出兼具荧光和金属信号的特异性分子探针用于荧光和质谱双模式成像。
市售的金属标签基于聚合物与金属离子螯合,金属担载量受金属配位效率和聚合度限制,而且聚合物配位的金属标签通过马来酰亚胺基团与IgG抗体上的四个巯基偶联,这对于低表达的标志物分子很难检测。基于纳米颗粒的金属标签大都存在较宽的尺寸范围,导致单颗粒上的金属含量区别较大,因此检测结果与生物标志物的表达高低不具有一致性。同时,纳米颗粒上金属含量过高可能造成仪器等离子体检测器的损坏或管道污染,还常常存在一定的非特异性吸附问题。已经报导过的荧光金属双信号纳米颗粒没有明确的荧光量子产率数据以及细胞爬片和组织切片染色以及用于荧光显微镜和质谱流式双模式成像的适用性验证。
因此,本领域的技术人员致力于开发一种具备荧光和金属双重信号的特异性探针用于免疫荧光和质谱流式双模式成像。
发明内容
有鉴于现有技术的上述缺陷,本发明所要解决的技术问题是提供可用于细胞爬片或组织切片上的生物标志物特异性标记,并具有荧光和金属双重信号,可用于双光子荧光显微镜成像和质谱流式成像技术的新型探针。
为实现上述目的,本发明提供了一种镧系金属掺杂碳量子点的制备方法,包括如下步骤:
将尿素和镧系金属盐Ln(NO3)3·nH2O,溶解于N,N-二甲基甲酰胺中,室温下磁力搅拌;
向上述溶液加入柠檬酸,溶解后转移至聚四氟乙烯内衬管内并放入不锈钢反应釜中;
将拧紧的所述不锈钢反应釜放入马弗炉反应后自然冷却至室温;
样品溶液转移至离心管内,离心,取上清液,得到沉淀物;
将所述沉淀物中加入乙醇,超声溶解,离心,取沉淀,重复上述离心直至上清液澄清;
将所述沉淀用去离子水溶解,用透析袋透析;
旋蒸浓缩,冻干得到黑色粉末,于干燥箱保存,得到CDs(Ln)。
本发明还提供一种基于上述的镧系金属掺杂碳量子点-核酸适配体偶联物的制备方法,其特征在于,包括如下步骤:
取所述CDs(Ln)与1-(3-二甲基氨基丙基)-3-乙基碳二胺盐酸盐和n-羟基琥珀酰亚胺混合,用无酶水稀释;孵育后,将活化的所述CDs(Ln)与3’-NH2修饰的抗前列腺膜抗原PSMA核酸适配体A10-3.2在室温下共价连接;最终用磷酸盐缓冲液PBS通过超滤管离心清洗未反应的碳点,得到产物CDs(Ln)-A10-3.2偶联物重悬于无酶水。
本发明还提供一种上述的CDs(Ln)-A10-3.2在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,包括如下步骤:
(1)细胞培养;
(2)细胞爬片;
(3)石蜡切片处理;
(4)细胞爬片及组织切片染色;
(5)质谱+荧光双成像技术对合成CDs(Ln)-A10-3.2纳米探针的靶向能力验证。
进一步地,步骤(1)中选择人前列腺癌LNCaP和PC-3细胞系分别为PSMA阳性和阴性表达细胞系,冻存细胞取出后37℃速溶,取15ml离心管,加入10ml含10%体积比胎牛血清的DMEM培养基,将冻存管中液体加入所述离心管中,轻吹混匀,1200转常温离心5min,去掉上清液,加入5ml含血清培养基,轻吹制成细胞悬液,加入到培养瓶中,后将所述培养瓶放置于37℃含50mL/L CO2的培养箱中进行细胞培养。
进一步地,步骤(2)中IMC仪器装载样本为载玻片,首先,爬片前用丙酮、乙醇和水对所述载玻片以及硅胶模具进行超声清洗,高压消毒灭菌,将所述硅胶模具按压到处理过的所述载玻片上,把细胞培养到对数生长期,消化后滴加到所述硅胶模具内,待1-2天后,细胞接近单层且贴壁完全后,吸取培养基,迅速用PBS漂洗2次,每次3-5秒,以清除血清;用新鲜配制的预冷的4%多聚甲醛缓冲液室温固定15min,PBS洗涤3遍后晾干保存于-20℃备用;可长期保存,做实验时,取出片子,PBS湿润后即可进行后续细胞染色实验。
进一步地,步骤(3)中首先对石蜡包埋的组织切片进行脱蜡和复水,将载玻片放入盛有预热的抗原修复溶液的离心管中95℃放置30min,抗原修复液以能没过组织所在的区域为准;30min后,将含有所述载玻片的离心管放置在实验室工作台上,自然冷却至室温;将所述载玻片转入盛有PBS的染缸,放置10分钟,洗涤,晾干,使用PAP笔在切片周围画圈。
进一步地,步骤(4)中用CDs(Ln)-A10-3.2探针对细胞爬片进行染色,主要考察不同的染色时间和探针浓度对结果的影响,通过共聚焦显微镜观察荧光标记情况,然后用成像质谱流式仪检测金属标记情况,对比二者的成像效果,优化染色条件。
进一步地,具体实验过程如下:在室温下用含有3%质量比BSA的DPBS对细胞封闭45min;吸去封闭溶液,将金属标记的探针分散液滴到细胞爬片的区域,4℃孵育过夜;用含有0.1%体积比的Triton-X的DPBS洗8min两次,放在摇床上缓慢摇晃;最后,在室温下用Ir-Intercalator对细胞爬片区域染色30min,用去离子水冲洗5min,室温晾干,待检测。
进一步地,步骤(5)中验证过程具体为:安捷伦Cary 60紫外-可见分光光度计和安捷伦Cary Eclipse荧光光谱仪测量镧系掺杂碳量子点的紫外吸收光谱和光致发光特性;采用AXIS Ultra DLD多功能X射线光电子能谱仪分析镧系金属掺杂碳点的X射线光电子能谱,其中包含了元素相对含量以及官能团信息;红外光谱特性采用EQUINOX 55型傅立叶红外光谱仪记录;细胞爬片的荧光图像采用蔡司LSM 880共聚焦显微镜采集;IMC图像通过Hyperion成像系统实现。
进一步地,所述Hyperion成像系统,激光类型:Nd:YAG,213nm;烧蚀斑大小:1μm2;135个通道,75-209amu。
总体而言,通过本发明所构思的以上技术方案与现有技术相比,主要具备以下的技术优点:镧系金属在生物体内含量极低,因而几乎没有背景信号;镧系金属掺杂碳量子点制备简单,在水溶液中金属和荧光信号稳定,而且成本低廉;镧系金属掺杂碳量子点尺寸小于10nm,分布均匀,水溶性好;核酸适配体对比抗体更稳定,成本更低,而且具备较好特异性;碳量子点表面富含羧基,容易与氨基修饰的核酸适配体偶联且保持探针良好的特异性;除检测金属元素外,只含有碳、氢、氧、氮,容易烧蚀,不会污染、堵塞或损坏仪器;核酸适配体包裹碳点的方式理论上一定程度避免了碳点的非特异性吸附,实验结果表明该探针在细胞爬片和肿瘤组织切片上均具备高特异性。
以下将结合附图对本发明的构思、具体结构及产生的技术效果作进一步说明,以充分地了解本发明的目的、特征和效果。
附图说明
图1是镧系金属掺杂碳量子点的制备过程示意图;
图2是CDs(Ho)的透射电镜形貌表征图;
图3是CDs(Ho)的紫外吸收光谱和荧光发射光谱图;
图4是CDs(Ho)和CDs(Tb)的XPS扫描全谱图;
图5是CDs(Ho)的XPS C1s分峰结果;
图6是CDs(Ho)的XPS O1s分峰结果;
图7是CDs(Ho)和CDs(Tb)的红外光谱图;
图8是CDs(Ho)-A10-3.2标染细胞爬片的荧光信号;
图9是CDs(Ho)-A10-3.2标染细胞爬片的165Ho质量信号的单细胞平均读数。
具体实施方式
以下参考说明书附图介绍本发明的多个优选实施例,使其技术内容更加清楚和便于理解。本发明可以通过许多不同形式的实施例来得以体现,本发明的保护范围并非仅限于文中提到的实施例。
实施例1:镧系金属掺杂碳量子点-核酸适配体偶联物的合成
1、溶剂热法制备镧系金属Ho,Tb掺杂碳量子点
将1g尿素和0.2g镧系金属盐(Tb(NO3)3·6H2O或Ho(NO3)3·6H2O),溶解于10mL N,N-二甲基甲酰胺中,室温下磁力搅拌1小时;
向上述溶液加入0.5g柠檬酸,溶解后转移至聚四氟乙烯内衬管内并放入不锈钢反应釜中;
将拧紧的反应釜放入马弗炉200℃反应4小时后自然冷却至室温;
样品溶液转移至离心管内,在20℃下以3000rpm的速度离心20min,取上清,得到沉淀物;
在沉淀中加入10mL乙醇,超声溶解,在4℃以15000rpm的速度离心40min,取沉淀重悬于乙醇。重复上述离心直至上清澄清。
将上述沉淀用去离子水溶解,用500kDa透析袋透析;
旋蒸浓缩,冻干得到黑色粉末,于干燥箱保存,得到CDs(Tb)和CDs(Ho)。
2、制备镧系金属Ho掺杂碳量子点-核酸适配体偶联物
取适量1mg/mL的CDs(Ho)与10μL,6.25mol/L的1-(3-二甲基氨基丙基)-3-乙基碳二胺盐酸盐(EDC)和10μL,6.25mol/L的n-羟基琥珀酰亚胺(NHS)混合,用无酶水(DNaseRNase-free water)稀释至80μL。37℃孵育30分钟后,将活化的Cdots(Ho)与20μL的3’-NH2修饰A10-3.2PSMA适配体(20μmol/L在无酶水中)在室温下共价连接4小时。最终用PBS缓冲液通过超滤管离心清洗未反应的碳点,得到产物CDs(Ho)-A10-3.2偶联物重悬于无酶水。
实施例2:CDs(Ho)-A10-3.2用于LNCaP细胞爬片的荧光显微成像和质谱流式成像。
(1)细胞培养:人前列腺癌LNCaP和PC-3细胞系分别为PSMA阳性和阴性表达细胞系。冻存细胞取出后37℃速溶,取15ml离心管,加入10ml含10%胎牛血清的DMEM培养基,将冻存管中液体(通常1.5ml)也加入离心管中,轻吹几下混匀,1200转常温离心5min,去掉上清,加入5ml含血清培养基,轻吹制成细胞悬液,加入到培养瓶中,后将培养瓶放置于37℃含50mL/L CO2的培养箱中进行细胞培养。
(2)细胞爬片:由于IMC仪器装载样本必须是载玻片,因此,我们采用特殊制作的硅胶模具让细胞单层生长在载玻片上。首先,爬片前用丙酮、乙醇和水对载玻片以及硅胶模具进行超声清洗,高压消毒灭菌(包括镊子和枪头)。将硅胶模具按压到处理过的载玻片上,把细胞培养到对数生长期,消化后滴加到硅胶模具内,待1-2天后,细胞接近单层且贴壁完全后(中间可加适量培养基),吸取培养基,迅速用PBS轻轻漂洗2次,每次3-5秒,以清除血清。用新鲜配制的预冷的4%多聚甲醛缓冲液室温固定15min,PBS(0.01mol/l,pH7.4)洗涤3遍后晾干保存于-20℃备用。可长期保存,做实验时,取出片子,入PBS湿润后即可进行后续细胞染色实验。
(3)石蜡切片处理:首先对石蜡包埋的组织切片进行脱蜡和复水,将带载玻片放入盛有预热的抗原修复溶液的离心管中95℃放置30min,抗原修复液以能没过组织所在的区域为准。30min后,将含有载玻片的离心管放置在实验室工作台上,自然冷却至室温,大概需要约30min的时间。将载玻片转入盛有PBS的染缸,放置10分钟,洗涤,晾干,使用PAP笔在切片周围画圈。
(4)细胞爬片及组织切片染色:用CDs(Ho)-A10-3.2探针对细胞爬片进行染色,以前列腺癌细胞系LNCaP为阳性细胞,PC-3为阴性细胞,主要考察不同的探针浓度对信号强度和特异性的影响,通过Confocal观察荧光标记情况,然后用IMC检测金属标记情况,对比二者的成像效果,优化染色条件。具体实验过程如下:在室温下用含有3%BSA的DPBS对细胞封闭45min。吸去封闭溶液,将金属标记的探针分散液滴到细胞爬片的区域,4℃孵育过夜。用含有0.1%Triton-X的DPBS洗8min两次,放在摇床上缓慢摇晃。最后,在室温下用Ir-Intercalator(201192B 1:400或者201192A 1:100DPBS混合液)对细胞爬片区域染色30min,用去离子水冲洗5min,室温晾干,待检测。质谱+荧光双成像技术对合成Blue-CDs(Ho)-A10-3.2探针的靶向能力进行了验证。
(5)我们使用TALOS F200X TALOS F200X场发射透射电镜在200千伏加速电压下,记录碳点形貌图像。镧系掺杂碳量子点的紫外吸收光谱和光致发光特性分别用安捷伦Cary60紫外-可见分光光度计和安捷伦Cary Eclipse荧光光谱仪测量。采用AXIS Ultra DLD多功能X射线光电子能谱仪(日本岛津Kratos公司)分析镧系金属掺杂碳点的X射线光电子能谱(XPS)。红外光谱采用EQUINOX 55型傅立叶红外光谱仪记录。荧光图像采用蔡司LSM 880共聚焦显微镜采集。IMC图像通过Hyperion成像系统(Fluidigm 108001);激光类型:Nd:YAG,213nm;烧蚀斑大小:1μm2;135个通道,75-209amu实现。收集的IMC图像通过MCD Viewer可视化,然后通过Cellprofiler软件分割单个细胞,最后用一个开源的计算组织学形态学细胞分析仪分析工具箱(histoCAT)软件提取单细胞数据,这些数据包括生物标志物的金属标记通道信号强度、细胞的大小和形状。
总之,本发明供一类可用于细胞爬片或组织切片上的生物标志物特异性标记,并具有荧光和金属双重信号,可用于双光子荧光显微镜成像和质谱流式成像技术的新型探针。对于用于细胞爬片及组织切片上生物标志物的特异性标记都需要探针具有良好的特异性。同时,分别以荧光显微镜和质谱流式成像作为互补的检测手段要求探针同时具备稳定的金属和荧光信号。
本发明提供一种可控合成镧系金属掺杂的多色荧光碳纳米颗粒CDs(Ln),通过与核酸适配体偶联制备了具备荧光金属双信号的标签探针,同时实现单细胞荧光质谱双模式成像。镧系金属在生物体内几乎没有背景信号,镧系金属掺杂的碳量子点具备较高的荧光量子产率和金属掺杂效率,同时与核酸适配体偶联后仍具备良好的靶向性。
首先通过溶剂热法制备掺杂一定量镧系金属的碳量子点,通过尿素与金属配位,然后以柠檬酸为碳源前驱体,N,N-二甲基甲酰胺为溶剂,高温下反应合成荧光金属标签CDs(Ln)。镧系金属性质相近,因而通过改变不同的镧系金属同位素掺杂就可以增加该金属标签的通道。
通过EDC-NHS方法偶联CDs(Ln)和氨基修饰的核酸适配体:上述合成的量子点表面富含羧基,通过EDC-NHS活化羧基,可通过与氨基修饰的适配体偶联。该方法耗时短,操作简单。
然后用上述步骤得到的探针(CDs(Ln)-Aptamer)负染细胞爬片或组织切片。首先,制作靶标分子阳性表达及阴性对照的细胞爬片,通过免疫荧光染色的方式用CDs(Ln)-Aptamer探针对细胞爬片进行染色,通过Confocal观察蓝色荧光的荧光强弱与靶标位置,然后用IMC检测金属标记情况,对比二者的成像效果,优化染色条件。然后对组织石蜡切片进行免疫荧光和IMC成像验证。
以上详细描述了本发明的较佳具体实施例。应当理解,本领域的普通技术无需创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此,凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验可以得到的技术方案,皆应在由权利要求书所确定的保护范围内。
Claims (8)
1.一种镧系金属掺杂碳量子点-核酸适配体偶联物CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,该应用是非疾病诊断和/或治疗目的,其特征在于,包括如下步骤:
(1)细胞培养;
(2)细胞爬片;
(3)石蜡切片处理;
(4)细胞爬片及组织切片染色;
(5)质谱+荧光双成像技术对合成CDs(Ln)- A10-3.2纳米探针的靶向能力验证;
所述CDs(Ln)- A10-3.2纳米探针的制备方法,包括如下步骤:
取CDs (Ln)与1-(3-二甲基氨基丙基)-3-乙基碳二胺盐酸盐和n-羟基琥珀酰亚胺混合,用无酶水稀释;孵育后,将活化的所述CDs(Ln)与3’- NH2修饰的抗前列腺膜抗原PSMA核酸适配体A10-3.2在室温下共价连接;最终用磷酸盐缓冲液PBS通过超滤管离心清洗未反应的碳点,得到产物CDs(Ln)- A10-3.2纳米探针重悬于无酶水;
所述CDs (Ln)的制备方法,包括如下步骤:
将尿素和镧系金属盐Ln(NO3)3·nH2O,溶解于N,N-二甲基甲酰胺中,室温下磁力搅拌;
向N,N-二甲基甲酰胺溶液中加入柠檬酸,溶解后转移至聚四氟乙烯内衬管内,并将聚四氟乙烯内衬管放入不锈钢反应釜中;
将拧紧的所述不锈钢反应釜放入马弗炉反应后自然冷却至室温;
样品溶液转移至离心管内,离心,取上清液,得到沉淀物;
在所述沉淀物中加入乙醇,超声溶解,离心,取沉淀,重悬于乙醇,重复上述离心直至上清液澄清;
将沉淀用去离子水溶解,用透析袋透析;
旋蒸浓缩,冻干得到黑色粉末,于干燥箱保存,得到CDs(Ln)。
2.如权利要求1所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,步骤(1)中选择人前列腺癌LNCaP和PC-3细胞系分别为PSMA阳性和阴性表达细胞系,冻存管取出后37℃速溶,取15ml离心管,加入10 ml含10%体积比胎牛血清的DMEM培养基,将冻存管中液体加入所述离心管中,轻吹混匀,1200转/min常温离心5min,去掉上清液,加入5 ml含血清培养基,轻吹制成细胞悬液,加入到培养瓶中,后将所述培养瓶放置于37℃含50 mL/L CO2的培养箱中进行细胞培养。
3.如权利要求1所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,步骤(2)中以载玻片为IMC仪器装载样本,首先,爬片前用丙酮、乙醇和水对所述载玻片以及硅胶模具进行超声清洗,高压消毒灭菌,将所述硅胶模具按压到处理过的所述载玻片上,把细胞培养到对数生长期,消化后滴加到所述硅胶模具内,待1-2天后,细胞接近单层且贴壁完全后,吸取培养基,迅速用PBS漂洗2次,每次3-5秒,以清除血清;用新鲜配制的预冷的4%多聚甲醛缓冲液室温固定15 min,PBS洗涤3遍后晾干保存于-20℃备用;可长期保存,做实验时,取出载玻片,PBS湿润后即可进行后续细胞染色实验。
4.如权利要求1所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,步骤(3)中首先对石蜡包埋的组织切片进行脱蜡和复水,将载玻片放入盛有预热的抗原修复溶液的离心管中95℃放置 30 min,抗原修复液以能没过组织所在的区域为准;30 min 后,将含有所述载玻片的离心管放置在实验室工作台上,自然冷却至室温;将所述载玻片转入盛有 PBS 的染缸,放置 10 分钟,洗涤,晾干,使用 PAP 笔在切片周围画圈。
5.如权利要求1所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,步骤(4)中用CDs(Ln)-A10-3.2纳米探针对细胞爬片进行染色,主要考察不同的染色时间和探针浓度对结果的影响,通过共聚焦显微镜观察荧光标记情况,然后用成像质谱流式仪检测金属标记情况,对比二者的成像效果,优化染色条件。
6.如权利要求1所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,具体实验过程如下:在室温下用含有 3%质量比 BSA的DPBS对细胞封闭45 min;吸去封闭溶液,将金属标记的CDs(Ln)- A10-3.2纳米探针分散液滴到细胞爬片的区域,4˚C 孵育过夜;用含有 0.1%体积比的Triton-X的DPBS洗8 min两次,放在摇床上缓慢摇晃;最后,在室温下用Ir-Intercalator对细胞爬片区域染色 30 min,用去离子水冲洗5 min,室温晾干,待检测。
7.如权利要求1所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,步骤(5)中验证过程具体为:安捷伦Cary 60紫外-可见分光光度计和安捷伦Cary Eclipse荧光光谱仪测量镧系金属掺杂碳量子点的紫外吸收光谱和光致发光特性;采用AXIS Ultra DLD多功能X射线光电子能谱仪分析镧系金属掺杂碳量子点的X射线光电子能谱,其中包含了元素相对含量以及官能团信息;红外光谱特性采用EQUINOX 55 型傅立叶红外光谱仪记录;细胞爬片的荧光图像采用蔡司LSM 880共聚焦显微镜采集;IMC图像通过Hyperion成像系统采集。
8.如权利要求7所述的CDs(Ln)- A10-3.2纳米探针在人前列腺癌细胞爬片和人前列腺癌组织切片的荧光显微成像和质谱流式成像中的应用,其特征在于,所述Hyperion成像系统,激光类型:Nd:YAG,213 nm;烧蚀斑大小:1μm2;135个通道,75-209 amu。
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