CN115667889A - 扫频荧光计 - Google Patents

扫频荧光计 Download PDF

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CN115667889A
CN115667889A CN202180036220.8A CN202180036220A CN115667889A CN 115667889 A CN115667889 A CN 115667889A CN 202180036220 A CN202180036220 A CN 202180036220A CN 115667889 A CN115667889 A CN 115667889A
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fluorescent species
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light source
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K·弗拉纳甘
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YSI Inc
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Abstract

扫频荧光计具有信号处理器或处理模块，该信号处理器或处理模块被配置为：接收信令，信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，液体样本被具有可变频率范围的光扫描，信息包括与液体中的感兴趣荧光物质相对应的特征光学频率，以及与感兴趣荧光物质保持在激发态的独特荧光寿命相对应的特征/寿命光学频率；以及基于所接收的信令，提供对应信令，该对应信令包含与感兴趣荧光物质的身份有关的信息，使用特征/寿命光学频率，感兴趣荧光物质被检测并且与液体中的重叠荧光物质区分开。

Description

扫频荧光计
相关申请的交叉引用
本申请要求于2020年5月20日提交的临时专利申请序列号63/027,587(911-023.9-1-1/N-YSI-0045US01)；于2020年5月21日提交的63/028,013号(911-023.010-1-1/N-YSI-0046US02)以及于2020年5月22日提交的63/028,723(911-023.011-1-1/N-YSI-0047US02)的权益，其全部内容通过引用并入本文。
技术领域
本发明涉及用于标识液体中的物质的荧光计，包括液体包含重叠物质的情况。
背景技术
通常，传统的荧光感测技术受到光谱宽广特征的困扰，尤其是当多个物质的荧光特征重叠时，该光谱宽广特征通常产生水物质的模糊标识。
具体地，传统的(单通道)荧光计典型地包括单个激发光源，单个激发光源被用于(在特定的光学波长下)光学激发感兴趣水参数、以及(在更长的光学波长下)重新发射感兴趣的水参数特定的光学光。这样的传感器通常利用锁定检测，该锁定检测选择性地测量固定(带内)频率的正弦信号，并拒绝具有其它频率(带外频率)的信号。
在本领域中需要更好的荧光计，例如用于标识包含重叠物质的液体中的物质。
发明内容
总之，根据本发明的传感器与传统荧光计的不同之处在于，针对该传感器的锁定频率将被“扫描”，即，从某个初始调制频率fi逐渐增加到某个最终调制频率ff。每个荧光物质具有其自身不同的荧光“寿命”(FLT)，在该寿命中，经激发的荧光团(由激发光学波长激发)在返回基态并伴随随后的发射之前在有限的时间(如寿命[秒])内保持在激发态。当信号的调制频率扫过物质的特征频率f＝(1/寿命)[Hz]时，除了信号振幅减小之外，信号的相位还经历90度的总体相对相移。相对相变的中点(45度)精确地对应于荧光物质的特征/寿命频率。简而言之，如果重叠荧光物质的身份存在不确定性，则寿命的知识可以进一步在两种物质之间进行描述。
实际上，(通过扫频激发频率的)荧光寿命保证了另一维的信息，增加了水体“指纹”的深度。
根据本发明的荧光计将测量：Ex/Em+用于提供增强检测技术的荧光寿命。
此外，本发明的一个关键在于沿着拟准直光的长度利用宽线性阵列的光机械配置。
本文的设计与浊度的、基于非强度的确定兼容。
例如，本发明可以应用于环境水质监测。
示例实施例
作为示例并且根据一些实施例，本发明可以包括或采取以信号处理器或处理模块为特征的装置，该信号处理器或处理模块被配置为：
接收信令，信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，液体样本被具有可变频率范围的光扫描，信息包括与液体中的感兴趣荧光物质相对应的特征光学频率，以及与独特荧光寿命相对应的特征/寿命光学频率，感兴趣荧光物质在独特荧光寿命中保持在激发态；以及
基于所接收的信令，提供对应信令，该对应信令包含与感兴趣荧光物质的身份有关的信息，使用特征/寿命光学频率，感兴趣荧光物质被检测并且与液体中的重叠荧光物质区分开。
装置可以包括以下附加特征中的一个或多个：
装置可以包括光传感器，该光传感器被配置为感测跨可变频率范围的光，并且提供由信号处理器或处理模块接收的信令。
光传感器可以是线性传感器阵列，其被配置为沿着提供光的光源的长度感测跨可变频率范围的光。
装置可以包括光源，该光源被配置为：跨可变频率范围，扫描具有激发光学波长的激发光，以激发一个或多个感兴趣荧光物质。
光源可以是扫频光源，该扫频光源包括拟准直光源。
可变频率范围可以包括锁定频率，该锁定频率通过将激发光从初始调制频率f1逐渐增加到最终调制频率ff而被扫描。
可变频率范围可以包括与特征/寿命光学频率相对应的中点频率。
装置可以包括扫频荧光计或采取扫频荧光计的形式。
扫频荧光计
作为进一步的示例，并且根据一些实施例，本发明可以采取以光源、光传感器和信号处理器或处理模块为特征的扫频荧光计的形式。
光源可以被配置为在具有一个或多个感兴趣荧光种类的液体样本上提供激发光，激发光具有可变频率范围，可变频率范围具有初始频率fi和最终频率ff。
光传感器可以被配置为感测从液体样本中的一个或多个感兴趣荧光物质反射的光，并且提供信令，该信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，该液体样本被具有可变频率范围的激发光扫描，该信息包括与液体中的感兴趣荧光物质相对应的特征光学频率，以及与感兴趣荧光物质保持在激发态的独特荧光寿命相对应的特征/寿命光学频率。
信号处理器或处理模块可以被配置为：
接收信令，以及
基于所接收的信令，提供对应信令，该对应信令包含与感兴趣荧光物质的身份有关的信息，使用特征/寿命光学频率，感兴趣荧光物质被检测并且与液体中的重叠荧光物质区分开。
方法
作为又一示例，并且根据一些实施例，本发明可以包括方法，方法的特征在于：
使用信号处理器或处理模块接收信令，信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，液体样本被具有可变频率范围的光扫描，信息包括与液体中的感兴趣荧光物质相对应的特征光学频率，以及与感兴趣荧光物质保持在激发态的独特荧光寿命相对应的特征/寿命光学频率；以及
使用信号处理器或处理模块基于所接收的信令，提供对应信令，该对应信令包含与感兴趣荧光物质的身份有关的信息，使用特征/寿命光学频率，感兴趣荧光物质被检测并且与液体中的重叠荧光物质区分开。
方法还可以包括上述特征中的一个或多个。
计算机可读存储介质
作为又一示例，并且根据本发明的一些实施例，本发明还可以采取计算机可读存储介质的形式，计算机可读存储介质具有用于执行前述方法的步骤的计算机可执行组件。计算机可读存储介质还可以包括上述特征中的一个或多个。
附图说明
不一定按比例绘制的附图包括图1-图5，如下：
图1是激发、发射和荧光寿命的曲线图，并且提供了根据本发明的荧光计的概念图，其中可以在相同的Ex/Em带通区域内测量三个不同但重叠的物质。传统的荧光计不具有区分三种物质的能力，但是根据本发明，利用荧光寿命的渲染，可以进行区分。
图2包括图2A和图2B，并且是两种重叠物质(TrisBipy Ru(II)和Pt Porphyrin)的激发发射矩阵(EEM)图。为了清楚起见，这些图被堆叠(不重叠)。两个帧中的轴和比例相同。
图3是相应物质(TrisBipy Ru(II)和Pt Porphyrin)的相对相位角(度)对扫频调制激发频率(Hz)的图或图线，例如，示出了相位响应如何完全可区分，但是它们相应的EEM标记占据了EEM图的相同区域，导致模糊。
图4是根据本发明的一些实施例的扫频荧光计的框图，该扫频荧光剂例如具有光源、光传感器和用于执行信号处理功能的信号处理器或处理模块。
图5是根据本发明的一些实施例的呈线性传感器阵列形式(例如，具有多行和多列光学元件)的光传感器的图。
为了减少附图中的混乱，附图中的每个图不必包括其中所示的每个元素的每个附图标记。
具体实施方式
图4示出了包括根据本发明的扫频荧光计的装置10，该装置10例如具有诸如拟准直光源的光源20、诸如线性传感器阵列的光传感器30以及信号处理器或处理模块40。
光源20可以被配置为：在具有一个或多个感兴趣荧光种类的液体样本上提供光，光具有可变频率范围，该可变频率范围具有初始频率fi和最终频率ff。根据一些实施例，光源20可以被配置为：对例如由信号处理器或处理模块40提供的光源控制信号做出响应，并且提供具有可变频率范围的光。
光传感器30可以被配置为：感测从液体样本中的感兴趣荧光物质反射的光；并且提供信令，该信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，该液体样本被具有可变频率范围的光扫描，该信息包括：特征光学频率，其与液体中的感兴趣荧光物质相对应；以及特征/寿命光学频率，其与感兴趣荧光物质保持在激发态的独特荧光寿命相对应。根据一些实施例，光传感器30可以被配置为：对例如由信号处理器或处理模块40提供的光传感器控制信号做出响应，并感测反射光。
信号处理器或处理模块40可以被配置为：
接收信令；以及
基于所接收的信令，提供对应信令，该对应信令包含与感兴趣荧光物质的身份有关的信息，该感兴趣荧光物质被检测、并且使用特征/寿命光学频率而与液体中的重叠荧光物质区分开。
根据一些实施例，信号处理器或处理模块40可以被配置为：例如向光源20和/或光传感器30提供光源控制信号、和/或光传感器控制信号。
光源20
作为示例，光源20可以被配置为：沿着线性传感器阵列30的对应长度(例如，如图5所示)提供光，光包括拟准直光，该拟准直光穿过相对于光源20和线性传感器阵列30布置的液体样本，以使得光从被监视或测试的液体样本中的一个或多个感兴趣荧光种类被反射到线性传感器阵列30上。
如本领域技术人员将理解的，拟准直光源在本领域中是已知的，并且本发明的范围不旨在限于现在已知或将来以后开发的任何特定类型或种类。
线性传感器阵列30
作为示例，线性传感器阵列30可以包括例如线性光电二极管阵列、线性电荷耦合器件(CCD)阵列或线性CMOS阵列。具体地，线性传感器阵列30可以包括类似于图5中所示的，例如是可单独寻址的光学元件的行和列的二维阵列(r1，c1；r1，c2；r1，c3；r1，c4；r1，c5；r1，c6；r1，c7；r1，c8；…；r1，cn；r2，c1；r2，c2；r2，c3；r2，c4；r2，c5；r2，c6；r2，c7；r2，c8；…；r2，cn；r3，c1；r3，c2；r3，c3；r3，c4；r3，c5；r3，c6；r3，c7；r3，c8；…；r3，cn；…；rn，c1；rn，c2；rn，c3；rn，c4；rn，c5；rn，c6；rn，c7；rn，c8；…；rn，cn)。线性传感器阵列在本领域中是已知的，并且本发明的范围不限于现在已知的或将来开发的任何特定类型或种类。
例如，在以下美国专利号9,020,202；8,022,349；7,956,341；7,040,538；5,252,818和4,193,057中公开了线性传感器阵列，其全部通过引用并入本文。
信号处理器或处理模块40
例如，信号处理器或处理模块40可以被配置为：基于如此确定的频率来确定一个或多个感兴趣荧光物质，然后基于该如此确定的频率来确定液体的浊度浓度。用于处理包含与所感测的光学频率有关的信息(例如，关于液体中的浊度浓度)的信令的技术在本领域中是已知的，并且本发明的范围不旨在限于现在已知的或将来以后开发的任何特定类型或种类。
信号处理功能的实现方式
例如，信号处理器或处理模块40的功能性可以使用硬件、软件、固件或其组合来实现。在典型的软件实现方式中，信号处理器40将包括基于一个或多个微处理器的架构，该架构具有例如至少一个信号处理器或微处理器。本领域的技术人员将能够利用适当的程序代码对诸如基于微控制器或基于微处理器的实现方式进行编程，以在无需过多实验的情况下，执行本文中所公开的信号处理功能。
本发明的范围不旨在限于使用现在已知的或将来以后开发的技术的任何特定实现方式。本发明的范围旨在包括将(多个)信号处理器的功能实现为独立处理器、信号处理器或信号处理器模块，以及单独处理器或处理器模块以及它们的某种组合。
作为示例，如本领域技术人员将理解的，装置10还可以包括例如整体由50指示的其他信号处理器电路或组件，包括随机存取存储器或存储器模块(RAM)和/或只读存储器(ROM)、输入/输出设备和控件以及将其连接的数据和地址总线和/或至少一个输入处理器和至少一个输出处理器。
作为另一示例，信号处理器可以包括或采取信号处理器和至少一个存储器的某种组合的形式，至少一个存储器包括计算机程序代码，其中信号处理器和至少一个存储器被配置为使得系统实现本发明的功能，例如，对所接收的信令做出响应，并基于所接收的信令来确定对应信令。
本发明的范围
虽然已参考示例性实施例描述了本发明，但是本领域技术人员将理解，在不脱离本发明的范围的情况下，可以进行各种改变，并且可以使用等同物来替代其元件。附加地，在不脱离本发明的基本范围的情况下，可以进行修改，以使得特定情况或材料适应本发明的教导。因此，本发明不限于本文中所公开的、作为实现本发明的最佳模式的(多个)特定实施例。

Claims (18)

1.一种装置，包括：
信号处理器或处理模块，其被配置为：
接收信令，所述信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，所述液体样本被具有可变频率范围的光扫描，所述信息包括与所述液体中的感兴趣荧光物质相对应的特征光学频率、以及与独特荧光寿命相对应的特征/寿命光学频率，所述感兴趣荧光物质在所述独特荧光寿命中保持在激发态；以及
基于所接收的所述信令，提供对应信令，所述对应信令包含与所述感兴趣荧光物质的身份有关的信息，使用所述特征/寿命光学频率，所述感兴趣荧光物质被检测并且与所述液体中的重叠荧光物质区分开。
2.根据权利要求1所述的装置，其中所述装置包括光传感器，所述光传感器被配置为感测跨所述可变频率范围的所述光，并提供所述信令。
3.根据权利要求2所述的装置，其中所述光传感器是线性传感器阵列，所述线性传感器阵列被配置为：沿着提供所述光的光源的长度，感测跨所述可变频率范围的所述光。
4.根据权利要求1所述的装置，其中所述装置包括光源，所述光源被配置为：跨所述可变频率范围，扫描具有激发光学波长的激发光，以激发所述一个或多个感兴趣荧光物质。
5.根据权利要求4所述的装置，其中所述光源是扫频光源，所述扫频光源包括拟准直光源。
6.根据权利要求1所述的装置，其中所述可变频率范围包括锁定频率，所述锁定频率通过将激发光从初始调制频率fi逐渐增加到最终调制频率ff而被扫描。
7.根据权利要求1所述的装置，其中所述可变频率范围包括与所述特征/寿命光学频率相对应的中点频率。
8.根据权利要求1所述的装置，其中所述装置是扫频荧光计。
9.一种方法，包括：
使用信号处理器或处理模块接收信令，所述信令包含与从液体样本中的一个或多个感兴趣荧光物质反射的光有关的信息，所述液体样本被具有可变频率范围的光扫描，所述信息包括与所述液体中的感兴趣荧光物质相对应的特征光学频率、以及与独特荧光寿命相对应的特征/寿命光学频率，所述感兴趣荧光物质在所述独特荧光寿命中保持在激发态；以及
使用所述信号处理器或处理模块基于所接收的所述信令、提供对应信令，所述对应信令包含与所述感兴趣荧光物质的身份有关的信息，使用所述特征/寿命光学频率，所述感兴趣荧光物质被检测并且与所述液体中的重叠荧光物质区分开。
10.根据权利要求9所述的方法，其中所述方法包括：将光传感器配置为感测跨所述可变频率范围的所述光，并提供所述信令。
11.根据权利要求10所述的方法，其中所述方法包括：将所述光传感器配置为线性传感器阵列，以沿着提供所述光的光源的长度，感测跨所述可变频率范围的所述光。
12.根据权利要求9所述的方法，其中所述方法包括：将光源配置为跨所述可变频率范围，扫描具有激发光学波长的激发光，以激发所述一个或多个感兴趣荧光物质。
13.根据权利要求12所述的方法，其中所述方法包括：将所述光源配置为扫频光源，所述扫频光源包括拟准直光源。
14.根据权利要求9所述的方法，其中所述可变频率范围包括锁定频率，所述锁定频率通过将激发光从初始调制频率fi逐渐增加到最终调制频率ff而被扫描。
15.根据权利要求9所述的方法，其中所述可变频率范围包括与所述特征/寿命光学频率相对应的中点频率。
16.根据权利要求9所述的方法，其中所述方法包括将所述信号处理器或信号处理模块配置为扫频荧光计的一部分。
17.一种扫频荧光计，包括：
光源，其被配置为在具有一个或多个感兴趣荧光物质的液体样本上提供光，所述光具有可变频率范围，所述可变频率范围具有初始频率fi和最终频率ff；
光传感器，其被配置为感测从所述液体样本中的所述感兴趣荧光物质反射的光，并且提供信令，所述信令包含与从所述液体样本中的所述一个或多个感兴趣荧光物质反射的光有关的信息，所述液体样本被具有可变频率范围的光扫描，所述信息包括与所述液体中的感兴趣荧光物质相对应的特征光学频率、以及与独特荧光寿命相对应的特征/寿命光学频率，所述感兴趣荧光物质在所述独特荧光寿命中保持在激发态；以及
信号处理器或处理模块，其被配置为：
接收所述信令，以及
基于所接收的所述信令，提供对应信令，所述对应信令包含与所述感兴趣荧光物质的身份有关的信息，使用特征/寿命光学频率，所述感兴趣荧光物质被检测并且与所述液体中的重叠荧光物质区分开。
18.根据权利要求17所述的扫频荧光计，其中
所述光源是扫频光源，所述扫频光源包括拟准直光源；并且
所述光传感器是线性传感器阵列，其被配置沿着提供所述光的光源的长度感测跨所述可变频率范围的所述光。
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