应用于生物芯片中的CMOS荧光检测系统的设计和研究
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摘要
基于荧光的检测方法是诸多生物实验中最重要的检测技术之一。而基于Si工艺的荧光检测芯片以其低成本,低功耗,高灵敏度等优点在很多荧光检测实验中获得了广泛的关注。本文着重研究一种基于CMOS工艺的荧光检测系统芯片的设计和荧光检测实验方案的改进,论文主要包括以下几方面内容:
在荧光检测系统设计方面,本文首先根据现有的工艺参数,建立了CMOS工艺光电二极管的光电转换模型,推导出光电转换效率的数学表达式,并且通过仿真和实际流片验证了该模型。除此,还分析比较了三种CMOS工艺兼容的光电二极管在灵敏度,峰值响应和暗电流等方面的差异,为荧光检测实验中选择适当的光电二极管提供了有利的理论基础。其次,本文提出一种新型的电容跨阻放大器(CTIA)结构读出电路来实现微弱荧光产生的光电流到电压的转换。这种结构拥有更高的光电流-电压转换增益,同时能够帮助光电二极管获得更好的暗电流特性。所提出的T型复位开关有效避免了积分期间复位开关漏电流的影响。最后,本文设计了一个12-bit,10-MS/s的流水线模数转换器(ADC)来实现检测芯片的数字化输出,为了获得较低的功耗和较小芯片面积,本文中的ADC采用了如下一些技术:通过对ADC模拟通路噪声和功耗模型的建立,确定2.5位的级分辨率;消除传统结构中的采样/保持电路;采用交换反馈电容开关(CFCS)技术来减轻级电路中采样电容的匹配要求;带有采样电容和预放大器的动态比较器;增益自举共源共栅放大器等等。本文通过SMIC 0.18-μm混合信号CMOS工艺对所设计的荧光检测芯片进行流片验证。芯片面积3 mm~2(包括PAD部分面积),消耗功耗37 mW。实验结果表明,所提出的光电转换模型与测试结果基本相符合,用于荧光检测实验的Nwell/Psub光电二极管在580 nm波长处拥有最大的光电转换灵敏度0.176 A/W。所采用的改进CTIA结构读出电路功能正确,电荷-电压转换增益为1.6μV/e~-,这种结构令Nwell/Psub光电二极管在300 mV反向偏压条件下,暗电流仅为300fA(3 nA/cm~2)。所设计的ADC的最大微分非线性(DNL)和最大积分非线性(INL)分别为+0.8 LSB和-3LSB。并且,当采样时钟为10 MHz,输入频率为1 MHz时,ADC与杂散无关的动态范围(SFDR)为61.7 dB,信号与噪声及谐波失真比(SNDR)为56.2 dB。
在荧光实验方面,本文采用了一种“接触式”检测方法,即将荧光产生池直接放到芯片表面,从而有效避免了额外的光学仪器和光学通路带来的荧光损失。实验证明,利用所设计的CMOS荧光检测芯片配合“接触式”检测方法能够在室温下成功检测到由荧光素溶液产生的荧光。当荧光素溶液体积为2 ml时,芯片在50 ms积分时间条件下,能够检测到的最小荧光强度为3.5 nW/cm~2,最小光电流7 fA,此时荧光素溶液浓度为20 ng/ml。当荧光素溶液体积为0.5μl时,芯片在10 ms积分时间内,能够检测到浓度为625 ng/ml的荧光素溶液产生的荧光,并在光电二极管内产生约为36 fA的光电流,此时照在芯片表面的荧光强度约为18 nW/cm~2。
Fluorescence-based method is one of the most important detecting techniques in various biological experiments.The fluorescent detecting chips based on Si technology attract widely concern in lots of detecting experiments because of their low cost,low power and high sensitivity.This thesis focuses on the study and design of a CMOS fluorescent detector chip,and the improvement of the fluorescent experiment scheme.The main contributions of this thesis are concluded as following:
On the design of the fluorescent detecting system,this thesis firstly establishes a photon-electron conversion model of the standard CMOS photodiodes based on some available technology parameters.The mathematic formulation of the conversion efficiency is also deduced and validated by simulation and testing.Furthermore,the comparisons of the three CMOS compatible photodiodes(N+/Psub photodiode,Nwell/Psub photodiode and P+/Nwell photodiode) on the sensitivity,peak response wavelength and dark current performance are given,which provide the theoretical basis for choosing proper photodectors in the detecting experiment.Secondly,a new capacitive trans-impedance amplifier(CTIA) is proposed to convert the photocurrent generated by weak fluorescence to voltage.This architecture has a higher photocurrent-voltage conversion gain and helps the photodiode have a better dark current performance.The proposed T-type reset switch avoids the leakage current influence effectively during the integrating period.Finally,a 12-bit, 10-MS/s pipelined analog-to-digital converter(ADC) is employed to realize the digitization outputs of the detecting chip.In order to achieve low power and small chip area,some special techniques have been adopted in this ADC.These techniques include the elimination of the conventional S/H circuit;the adoption of 2.5-bit/stage according to the noise model and power model in analog chain;the commutated feedback-capacitor switching(CFCS) technique to release the sampling capacitor mismatch requirement in the stage circuit; dynamic comparators with sampling capacitors and pre-amplifiers;gain boosting cascode amplifiers and so on.The proposed fluorescent detecting system is implemented in SMIC 0.18-μm standard CMOS process.This chip occupies an area of 3 mm~2(including PAD area) and consumes 37 roW.Experiments show that,the proposed photon-electron conversion model of the CMOS compatible photodiodes identifies the testing results.The Nwell/Psub photodiode used in the fluorescence detecting experiment has the maximum sensitivity of 0.176 A/W at wavelength of 580 nm.The CTIA has a charge-voltage conversion gain of 1.6μV/e~- and helps the Nwell/Psub photodiode to achieve a dark current of 300 fA(3 nA/cm~2) with 300 mV reverse biased voltage.The maximum DNL and INL of the designed ADC are +0.8 LSB and -3 LSB,respectively.A signal to noise and distortion ratio(SNDR) of 56.2 dB and a spurious free dynamic range(SFDR) of 61.7 dB are achieved at 1 MHz input frequency and 10 MHz sampling rate.
On the testing method of the fluorescence detecting experiment,a "contact imaging" detecting method,which puts the reaction vessel onto the surface of the detecting chip directly,is adopted to avoid the fluorescence loss due to the additional optical instruments and paths.Testing results indicate that,the detector system cooperating with the "contact imaging" detecting method can detect the fluorescence generated by fluorescein solution successfully at room temperature.With a solution volume of 2 ml and an integrating time of 50 ms,the detecting chip is sensitive to the fluorescence(3.5 nW/cm~2) excited from the fluorescein solution with a concentration of 20 ng/ml,and can generate a photocurrent of 7 fA within the photodetector.While with a solution volume of 0.5μl and an integrating time of 10 ms,the detecting chip is also sensitive to the fluorescence(18 nW/cm~2) excited from the fluorescein solution with a concentration of 625 ng/ml,and generates a photocurrent of 36fA.
在荧光检测系统设计方面,本文首先根据现有的工艺参数,建立了CMOS工艺光电二极管的光电转换模型,推导出光电转换效率的数学表达式,并且通过仿真和实际流片验证了该模型。除此,还分析比较了三种CMOS工艺兼容的光电二极管在灵敏度,峰值响应和暗电流等方面的差异,为荧光检测实验中选择适当的光电二极管提供了有利的理论基础。其次,本文提出一种新型的电容跨阻放大器(CTIA)结构读出电路来实现微弱荧光产生的光电流到电压的转换。这种结构拥有更高的光电流-电压转换增益,同时能够帮助光电二极管获得更好的暗电流特性。所提出的T型复位开关有效避免了积分期间复位开关漏电流的影响。最后,本文设计了一个12-bit,10-MS/s的流水线模数转换器(ADC)来实现检测芯片的数字化输出,为了获得较低的功耗和较小芯片面积,本文中的ADC采用了如下一些技术:通过对ADC模拟通路噪声和功耗模型的建立,确定2.5位的级分辨率;消除传统结构中的采样/保持电路;采用交换反馈电容开关(CFCS)技术来减轻级电路中采样电容的匹配要求;带有采样电容和预放大器的动态比较器;增益自举共源共栅放大器等等。本文通过SMIC 0.18-μm混合信号CMOS工艺对所设计的荧光检测芯片进行流片验证。芯片面积3 mm~2(包括PAD部分面积),消耗功耗37 mW。实验结果表明,所提出的光电转换模型与测试结果基本相符合,用于荧光检测实验的Nwell/Psub光电二极管在580 nm波长处拥有最大的光电转换灵敏度0.176 A/W。所采用的改进CTIA结构读出电路功能正确,电荷-电压转换增益为1.6μV/e~-,这种结构令Nwell/Psub光电二极管在300 mV反向偏压条件下,暗电流仅为300fA(3 nA/cm~2)。所设计的ADC的最大微分非线性(DNL)和最大积分非线性(INL)分别为+0.8 LSB和-3LSB。并且,当采样时钟为10 MHz,输入频率为1 MHz时,ADC与杂散无关的动态范围(SFDR)为61.7 dB,信号与噪声及谐波失真比(SNDR)为56.2 dB。
在荧光实验方面,本文采用了一种“接触式”检测方法,即将荧光产生池直接放到芯片表面,从而有效避免了额外的光学仪器和光学通路带来的荧光损失。实验证明,利用所设计的CMOS荧光检测芯片配合“接触式”检测方法能够在室温下成功检测到由荧光素溶液产生的荧光。当荧光素溶液体积为2 ml时,芯片在50 ms积分时间条件下,能够检测到的最小荧光强度为3.5 nW/cm~2,最小光电流7 fA,此时荧光素溶液浓度为20 ng/ml。当荧光素溶液体积为0.5μl时,芯片在10 ms积分时间内,能够检测到浓度为625 ng/ml的荧光素溶液产生的荧光,并在光电二极管内产生约为36 fA的光电流,此时照在芯片表面的荧光强度约为18 nW/cm~2。
Fluorescence-based method is one of the most important detecting techniques in various biological experiments.The fluorescent detecting chips based on Si technology attract widely concern in lots of detecting experiments because of their low cost,low power and high sensitivity.This thesis focuses on the study and design of a CMOS fluorescent detector chip,and the improvement of the fluorescent experiment scheme.The main contributions of this thesis are concluded as following:
On the design of the fluorescent detecting system,this thesis firstly establishes a photon-electron conversion model of the standard CMOS photodiodes based on some available technology parameters.The mathematic formulation of the conversion efficiency is also deduced and validated by simulation and testing.Furthermore,the comparisons of the three CMOS compatible photodiodes(N+/Psub photodiode,Nwell/Psub photodiode and P+/Nwell photodiode) on the sensitivity,peak response wavelength and dark current performance are given,which provide the theoretical basis for choosing proper photodectors in the detecting experiment.Secondly,a new capacitive trans-impedance amplifier(CTIA) is proposed to convert the photocurrent generated by weak fluorescence to voltage.This architecture has a higher photocurrent-voltage conversion gain and helps the photodiode have a better dark current performance.The proposed T-type reset switch avoids the leakage current influence effectively during the integrating period.Finally,a 12-bit, 10-MS/s pipelined analog-to-digital converter(ADC) is employed to realize the digitization outputs of the detecting chip.In order to achieve low power and small chip area,some special techniques have been adopted in this ADC.These techniques include the elimination of the conventional S/H circuit;the adoption of 2.5-bit/stage according to the noise model and power model in analog chain;the commutated feedback-capacitor switching(CFCS) technique to release the sampling capacitor mismatch requirement in the stage circuit; dynamic comparators with sampling capacitors and pre-amplifiers;gain boosting cascode amplifiers and so on.The proposed fluorescent detecting system is implemented in SMIC 0.18-μm standard CMOS process.This chip occupies an area of 3 mm~2(including PAD area) and consumes 37 roW.Experiments show that,the proposed photon-electron conversion model of the CMOS compatible photodiodes identifies the testing results.The Nwell/Psub photodiode used in the fluorescence detecting experiment has the maximum sensitivity of 0.176 A/W at wavelength of 580 nm.The CTIA has a charge-voltage conversion gain of 1.6μV/e~- and helps the Nwell/Psub photodiode to achieve a dark current of 300 fA(3 nA/cm~2) with 300 mV reverse biased voltage.The maximum DNL and INL of the designed ADC are +0.8 LSB and -3 LSB,respectively.A signal to noise and distortion ratio(SNDR) of 56.2 dB and a spurious free dynamic range(SFDR) of 61.7 dB are achieved at 1 MHz input frequency and 10 MHz sampling rate.
On the testing method of the fluorescence detecting experiment,a "contact imaging" detecting method,which puts the reaction vessel onto the surface of the detecting chip directly,is adopted to avoid the fluorescence loss due to the additional optical instruments and paths.Testing results indicate that,the detector system cooperating with the "contact imaging" detecting method can detect the fluorescence generated by fluorescein solution successfully at room temperature.With a solution volume of 2 ml and an integrating time of 50 ms,the detecting chip is sensitive to the fluorescence(3.5 nW/cm~2) excited from the fluorescein solution with a concentration of 20 ng/ml,and can generate a photocurrent of 7 fA within the photodetector.While with a solution volume of 0.5μl and an integrating time of 10 ms,the detecting chip is also sensitive to the fluorescence(18 nW/cm~2) excited from the fluorescein solution with a concentration of 625 ng/ml,and generates a photocurrent of 36fA.
引文
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