CN114471760A - 一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置及使用方法 - Google Patents
一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置及使用方法 Download PDFInfo
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Abstract
本发明公开了一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置及使用方法。所述微流体芯片包括样本通道、两个鞘液通道、第一荧光检测区、第二荧光检测区、磁场控制***、磁场控制细胞分选区、靶细胞通道以及废液通道;基于细胞上标记的荧光信号,将FACS***与磁场控制分选***相结合,实现了自动化细胞分选,不仅将分选检测集成小型化和自动化、而且具有操作简易、经济等优点。该装置分选既可以分选大量细胞样本,也可以分选少量细胞样本,克服了流式细胞仪的缺陷。
Description
技术领域
本发明涉及一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置及使用方法,属于微流体芯片领域。
背景技术
流式细胞仪(Flow Cytometer)是一种在功能水平上对单细胞或其他生物粒子进行定量分析和分选的检测手段,分析速度可达上万个细胞/秒,同时能从一个细胞中测得多个参数。细胞分选也是它的重要应用之一。它能够根据每个细胞的光散射和荧光特征,将含特定细胞的液滴赋予电荷,并在下游集成高压电场。通过细胞信号的反馈控制高压电场,使含有细胞的液滴受电场力发生偏移,最终收集到目标容器中。故人们又将流式细胞仪称作荧光激活细胞分选仪(Fluorescence-activated Cell Sorter,FACS)。FACS可高纯度识别、分离细胞群,特别是稀有细胞群体。在分离纯化表型已知特定细胞群的各种方法中,FACS脱颖而出,适用于往往需要高纯度细胞群的实验和临床研究。但是,FACS不仅设备庞大、昂贵,而且分析成本高、单次分析需要大量的检测样本。
发明内容
本发明提供了一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置及使用方法,基于细胞上标记的荧光信号,将FACS***与磁场控制分选***相结合,实现了自动化细胞分选,不仅将分选检测集成小型化和自动化、而且具有操作简易、经济等优点。该装置分选既可以分选大量细胞样本,也可以分选少量细胞样本,克服了流式细胞仪的缺陷。
本发明采用以下技术方案:
一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置,包括样本通道、两个鞘液通道、第一荧光检测区、第二荧光检测区、磁场控制***、磁场控制细胞分选区、靶细胞通道以及废液通道;
所述样本通道和两个鞘液通道相连通,样本通道和两个鞘液通道互相并联,样本通道位于两个鞘液通道之间;所述样本通道与两个鞘液通道的连接管道分别为第一流道和第二流道;第一流道和第二流道与两个鞘液通道分别交叉于第一交叉点和第二交叉点;所述靶细胞通道始于所述第一交叉点,废液通道始于所述第二交叉点;所述磁场控制细胞分选区设置在第一流道和第二流道上,所述细胞分选区包括一个磁子;所述磁子由磁场控制***控制,在第一流道和第二流道上来回移动,以控制样本的流向;当磁子位于第一流道时,样本通道与废液通道相连通,当磁子位于第二流道时,样本通道与靶细胞通道相连通;
所述第一荧光检测区位于样本通道上;第二荧光检测区位于靶细胞通道上;
所述样本通道的进样口和两个鞘液通道的进样口分别连接注射器与相应注射泵,为溶液在芯片内流动提供动力;
所述废液通道的出样口与废液专用收集瓶相连;用于收集非分选所需的细胞以及多余的细胞废液;
所述靶细胞通道的出样口连接靶细胞专用瓶,用于收集含有目标细胞的溶液。
进一步地,所述样本通道进样口连接的注射器内为预处理的细胞样本溶液;所述两个鞘液通道进样口连接的注射器内为相应的鞘液溶液、细胞缓冲液或培养液。
进一步地,所述第一荧光检测区和第二荧光检测区结构相同,包括激光和荧光检测器,所述激光和荧光检测器分别位于芯片的两侧;所述荧光检测器含有滤光片。
进一步地,所述滤光片能够将非荧光信号滤掉,只保留荧光进入检测器。
进一步地,所述第一流道和第二流道上在第一交叉点和第二交叉点处分别设有一个限位凹槽,用于使磁子封堵通往靶细胞通道或废液通道的路径。
进一步地,所述芯片装置由化学惰性、光学透明、生物相容性的塑料材质制成。
进一步地,所述芯片装置的材质包括聚二甲基硅氧烷、PMMA、COC、COP。
所述基于磁场控制分选荧光标记细胞方法的微流体芯片装置的使用方法,包括以下步骤:
S1、准备进样时,首先开启激光、检测器以及磁场控制***,然后开启三个进样注射泵;
S2、进样时,样本通道进样口内注入的溶液为荧光抗体孵育过的预处理细胞样本溶液,同时鞘液通道进样口内注入的溶液为鞘液溶液;
S3、预处理样本溶液流经第一荧光检测区和第二荧光检测区时,激光和检测器会依次对溶液内进行检测,检测结果经计算机***处理并反馈给计算机***控制的磁场控制***,磁场控制***则会控制磁场中的磁子的移动方向;
S31、若第一荧光检测区未检测到具有荧光标记信号的细胞,磁场控制***则会控制磁子移动至第一流道并封堵通往靶细胞通道的路径,使样本溶液向废液通道的出样口流动,最终在废液专用瓶中收集;
S32、若第一荧光检测区检测到具有荧光标记信号的细胞,磁场控制***则会控制磁子移动至第二流道并封堵通往废液通道的路径,使含有目标细胞的样本溶液向靶细胞通道的出样口流动,并且流经第二荧光检测区,二次确认是否存在第一荧光检测区所检测到的目标细胞,若二次确认检测到目标细胞,***控制磁子回到第一流道并封堵通往靶细胞通道的路径;
S33、重复步骤S31和步骤S32,并最终在靶细胞专用瓶中收集获得目标细胞。
有益效果
(1)相对于传统的流式荧光细胞分选手段,该装备将分选检测集成小型化和自动化,操作简易、经济。
(2)不受细胞样本量的控制,既可以大样本也可以小样本。
(3)对样本进行预处理之后,所有操作均在芯片内进行,污染小。
(4)细胞分选发生在芯片管道内部,分选准确度更高。
附图说明
图1.装置立体结构设计图。
图2.细胞分选区结构图;图2a为细胞分选区上游荧光检测区未检测到荧光信号的情况;图2b为细胞分选区上游荧光检测区检测到有荧光信号的情况。
图3.荧光检测区分布结构图。
图4.荧光检测区侧面图。
具体实施方式
下面详细描述本发明的实施方式,所述实施方式的示例在附图中示出。下面通过参考附图描述的实施方式是示例性的,仅用于解释本发明,而不能解释为对本发明的限制。
如图1和图2所示,一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置,包括样本通道、两个鞘液通道、第一荧光检测区、第二荧光检测区、磁场控制***、磁场控制细胞分选区、靶细胞通道以及废液通道;所述芯片装置由化学惰性、光学透明、生物相容性的塑料材质制成;所述芯片装置的材质包括聚二甲基硅氧烷、PMMA、COC、COP。
所述样本通道和两个鞘液通道相连通,样本通道和两个鞘液通道互相并联,样本通道位于两个鞘液通道之间;所述样本通道与两个鞘液通道的连接管道分别为第一流道和第二流道;第一流道和第二流道与两个鞘液通道分别交叉于第一交叉点和第二交叉点;所述靶细胞通道始于所述第一交叉点,废液通道始于所述第二交叉点;所述磁场控制细胞分选区设置在第一流道和第二流道上,所述细胞分选区包括一个磁子;所述磁子由磁场控制***控制,在第一流道和第二流道上来回移动,以控制样本的流向;当磁子位于第一流道时,样本通道与废液通道相连通,当磁子位于第二流道时,样本通道与靶细胞通道相连通;所述第一流道和第二流道上在第一交叉点和第二交叉点处分别设有一个限位凹槽,用于使磁子封堵通往靶细胞通道或废液通道的路径。
如图3和图4所示,所述第一荧光检测区位于样本通道上;第二荧光检测区位于靶细胞通道上;所述第一荧光检测区和第二荧光检测区结构相同,包括激光和荧光检测器,所述激光和荧光检测器分别位于芯片的两侧;所述荧光检测器含有滤光片。位于荧光检测区下方的激光器发射激光光源(虚线方框),并持续照射在荧光检测区,最终光信号经过荧光检测区上方的滤光片和激光检测器。当荧光检测区存在荧光信号时,滤光片可将非荧光信号过滤掉,使激光检测器只捕捉到荧光信号;当荧光检测区无荧光信号存在时,经激光照射,并通过滤光片后,激光检测器内无信号计入。
所述样本通道的进样口和两个鞘液通道的进样口分别连接注射器与相应注射泵,为溶液在芯片内流动提供动力;所述样本通道进样口连接的注射器内为预处理的细胞样本溶液;所述两个鞘液通道进样口连接的注射器内为相应的鞘液溶液、细胞缓冲液或培养液。所述废液通道的出样口与废液专用收集瓶相连;用于收集非分选所需的细胞以及多余的细胞废液;所述靶细胞通道的出样口连接靶细胞专用瓶,用于收集含有目标细胞的溶液。
所述基于磁场控制分选荧光标记细胞方法的微流体芯片装置的使用方法,包括以下步骤:
S1、准备进样时,首先开启激光、检测器以及磁场控制***,然后开启三个进样注射泵;
S2、进样:样本通道进样口内注入的溶液为荧光抗体孵育过的预处理细胞样本溶液,同时鞘液通道进样口内注入的溶液为鞘液溶液;
S3、预处理样本溶液流经第一荧光检测区和第二荧光检测区时,激光和检测器会依次对溶液行检测,检测结果经计算机***处理并反馈给计算机***控制的磁场控制***,磁场控制***则会控制磁场中的磁子的移动方向;
S31、若第一荧光检测区未检测到具有荧光标记信号的细胞,磁场控制***则会控制磁子移动至第一流道并封堵通往靶细胞通道的路径,使样本溶液向废液通道的出样口流动,最终在废液专用瓶中收集;
S32、若第一荧光检测区检测到具有荧光标记信号的细胞,磁场控制***则会控制磁子移动至第二流道并封堵通往废液通道的路径,使含有目标细胞的样本溶液向靶细胞通道的出样口流动,并且流经第二荧光检测区,二次确认是否存在第一荧光检测区所检测到的目标细胞,若二次确认检测到目标细胞,***控制磁子回到第一流道并封堵通往靶细胞通道的路径;
S33、重复步骤S31和步骤S32,并最终在靶细胞专用瓶中收集获得目标细胞。
Claims (8)
1.一种基于磁场控制分选荧光标记细胞方法的微流体芯片装置,其特征在于:所述微流控芯片包括样本通道、两个鞘液通道、第一荧光检测区、第二荧光检测区、磁场控制***、磁场控制细胞分选区、靶细胞通道以及废液通道;
其中,所述样本通道和两个鞘液通道相连通,样本通道和两个鞘液通道互相并联,样本通道位于两个鞘液通道之间;所述样本通道与两个鞘液通道的连接管道分别为第一流道和第二流道;第一流道和第二流道与两个鞘液通道分别交叉于第一交叉点和第二交叉点;所述靶细胞通道始于所述第一交叉点,废液通道始于所述第二交叉点;
所述磁场控制细胞分选区设置在第一流道和第二流道上,所述细胞分选区包括一个磁子;所述磁子由磁场控制***控制,在第一流道和第二流道上来回移动,以控制样本的流向;当磁子位于第一流道时,样本通道与废液通道相连通,当磁子位于第二流道时,样本通道与靶细胞通道相连通;
所述第一荧光检测区位于样本通道上;第二荧光检测区位于靶细胞通道上;
所述样本通道的进样口和两个鞘液通道的进样口分别连接注射器与相应注射泵,为溶液在芯片内流动提供动力;
所述废液通道的出样口与废液专用收集瓶相连;用于收集非分选所需的细胞以及多余的细胞废液;
所述靶细胞通道的出样口连接靶细胞专用瓶,用于收集含有目标细胞的溶液。
2.根据权利要求1所述装置,其特征在于,所述样本通道进样口连接的注射器内为预处理的细胞样本溶液;所述两个鞘液通道进样口连接的注射器内为相应的鞘液溶液、细胞缓冲液或培养液。
3.根据权利要求1所述装置,其特征在于,所述第一荧光检测区和第二荧光检测区结构相同,包括激光和荧光检测器,所述激光和荧光检测器分别位于芯片的两侧;所述荧光检测器含有滤光片。
4.根据权利要求3所述装置,其特征在于,所述滤光片能够将非荧光信号滤掉,只保留荧光进入检测器。
5.根据权利要求1所述装置,其特征在于,所述第一流道和第二流道上在第一交叉点和第二交叉点处分别设有一个限位凹槽,用于使磁子封堵通往靶细胞通道或废液通道的路径。
6.根据权利要求1所述装置,其特征在于,所述芯片装置由化学惰性、光学透明、生物相容性的塑料材质制成。
7.根据权利要求6所述装置,其特征在于,所述芯片装置的材质包括聚二甲基硅氧烷、PMMA、COC、COP。
8.根据权利要求1-7任一项所述的基于磁场控制分选荧光标记细胞方法的微流体芯片装置的使用方法,其特征在于,所述方法包括以下步骤:
S1、准备进样时,首先开启激光、检测器以及磁场控制***,然后开启三个进样注射泵;
S2、进样:样本通道进样口内注入的溶液为荧光抗体孵育过的预处理细胞样本溶液,同时鞘液通道进样口内注入的溶液为鞘液溶液;
S3、预处理样本溶液流经第一荧光检测区和第二荧光检测区时,激光和检测器会依次对溶液内进行检测,检测结果经计算机***处理并反馈给计算机***控制的磁场控制***,磁场控制***则会控制磁场中的磁子的移动方向;
S31、若第一荧光检测区未检测到具有荧光标记信号的细胞,磁场控制***则会控制磁子移动至第一流道并封堵通往靶细胞通道的路径,使样本溶液向废液通道的出样口流动,最终在废液专用瓶中收集;
S32、若第一荧光检测区检测到具有荧光标记信号的细胞,磁场控制***则会控制磁子移动至第二流道并封堵通往废液通道的路径,使含有目标细胞的样本溶液向靶细胞通道的出样口流动,并且流经第二荧光检测区,二次确认是否存在第一荧光检测区所检测到的目标细胞,若二次确认检测到目标细胞,***控制磁子回到第一流道并封堵通往靶细胞通道的路径;
S33、重复步骤S31和步骤S32,并最终在靶细胞专用瓶中收集获得目标细胞。
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Publication number | Priority date | Publication date | Assignee | Title |
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CN116337728A (zh) * | 2023-05-30 | 2023-06-27 | 天津大学 | 一种单片集成微流式细胞仪荧光检测装置 |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070178529A1 (en) * | 2006-01-13 | 2007-08-02 | Micronics, Inc. | Electromagnetically actuated valves for use in microfluidic structures |
CN104511324A (zh) * | 2013-10-01 | 2015-04-15 | Owl生物医学公司 | 带有面外通道和会聚元件的颗粒操纵*** |
CN104830664A (zh) * | 2015-05-07 | 2015-08-12 | 清华大学 | 一种基于外置压电陶瓷驱动的微流体细胞分选*** |
CN104877898A (zh) * | 2014-02-27 | 2015-09-02 | 中国科学院青岛生物能源与过程研究所 | 一种低成本、高效分离获取单细胞的***和方法 |
CN106999927A (zh) * | 2014-09-30 | 2017-08-01 | 福斯分析仪器公司 | 用于流体动力流动聚焦的方法、装置和*** |
CN108778509A (zh) * | 2016-03-08 | 2018-11-09 | 生物辐射实验室股份有限公司 | 微流体颗粒分选器 |
CN109943475A (zh) * | 2019-04-12 | 2019-06-28 | 广西医科大学第一附属医院 | 一种微流控分选芯片及其分选*** |
CN111295578A (zh) * | 2017-08-15 | 2020-06-16 | 华盛顿大学 | 颗粒分离***和方法 |
CN113188980A (zh) * | 2021-04-28 | 2021-07-30 | 南通大学 | 一种基于荧光激活细胞分选技术的全血循环肿瘤细胞级联分选的装置及方法 |
-
2022
- 2022-02-10 CN CN202210125750.6A patent/CN114471760A/zh active Pending
- 2022-09-01 US US17/929,099 patent/US20230249182A1/en active Pending
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070178529A1 (en) * | 2006-01-13 | 2007-08-02 | Micronics, Inc. | Electromagnetically actuated valves for use in microfluidic structures |
CN104511324A (zh) * | 2013-10-01 | 2015-04-15 | Owl生物医学公司 | 带有面外通道和会聚元件的颗粒操纵*** |
CN104877898A (zh) * | 2014-02-27 | 2015-09-02 | 中国科学院青岛生物能源与过程研究所 | 一种低成本、高效分离获取单细胞的***和方法 |
CN106999927A (zh) * | 2014-09-30 | 2017-08-01 | 福斯分析仪器公司 | 用于流体动力流动聚焦的方法、装置和*** |
CN104830664A (zh) * | 2015-05-07 | 2015-08-12 | 清华大学 | 一种基于外置压电陶瓷驱动的微流体细胞分选*** |
CN108778509A (zh) * | 2016-03-08 | 2018-11-09 | 生物辐射实验室股份有限公司 | 微流体颗粒分选器 |
CN111295578A (zh) * | 2017-08-15 | 2020-06-16 | 华盛顿大学 | 颗粒分离***和方法 |
CN109943475A (zh) * | 2019-04-12 | 2019-06-28 | 广西医科大学第一附属医院 | 一种微流控分选芯片及其分选*** |
CN113188980A (zh) * | 2021-04-28 | 2021-07-30 | 南通大学 | 一种基于荧光激活细胞分选技术的全血循环肿瘤细胞级联分选的装置及方法 |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116337728A (zh) * | 2023-05-30 | 2023-06-27 | 天津大学 | 一种单片集成微流式细胞仪荧光检测装置 |
CN116337728B (zh) * | 2023-05-30 | 2023-08-18 | 天津大学 | 一种单片集成微流式细胞仪荧光检测装置 |
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