Author Affiliations
Abstract
1 Institute of Semiconductors, Chinese Academy of Sciences, Beijing 100083, China
2 College of Materials Science and Opto-Electronic Technology, University of Chinese Academy of Sciences, Beijing 100049, China
3 Beijing Key Laboratory of Genome and Precision Medicine Technologies, Beijing 100101, China
Ion sensitive field effect transistor (ISFET) devices are highly accurate, convenient, fast and low-cost in the detection of ions and biological macromolecules, such as DNA molecules, antibodies, enzymatic substrates and cellular metabolites. For high-throughput cell metabolism detection, we successfully designed a very large-scale biomedical sensing application specific integrated circuit (ASIC) with a 640 × 640 ISFET array. The circuit design is highly integrated by compressing the size of a pixel to 7.4 × 7.4μm2 and arranging the layout of even and odd columns in an interdigital pattern to maximize the utilization of space. The chip can operate at a speed of 2.083M pixels/s and the dynamic process of the fluid flow on the surface of the array was monitored through ion imaging. The pH sensitivity is 33 ± 4 mV/pH and the drift rate is 0.06 mV/min after 5 h, indicating the stability and robustness of the chip. Moreover, the chip was applied to monitor pH changes in CaSki cells metabolism, with pH shifting from 8.04 to 7.40 on average. This platform has the potential for continuous and parallel monitoring of cell metabolism in single-cell culture arrays.Ion sensitive field effect transistor (ISFET) devices are highly accurate, convenient, fast and low-cost in the detection of ions and biological macromolecules, such as DNA molecules, antibodies, enzymatic substrates and cellular metabolites. For high-throughput cell metabolism detection, we successfully designed a very large-scale biomedical sensing application specific integrated circuit (ASIC) with a 640 × 640 ISFET array. The circuit design is highly integrated by compressing the size of a pixel to 7.4 × 7.4μm2 and arranging the layout of even and odd columns in an interdigital pattern to maximize the utilization of space. The chip can operate at a speed of 2.083M pixels/s and the dynamic process of the fluid flow on the surface of the array was monitored through ion imaging. The pH sensitivity is 33 ± 4 mV/pH and the drift rate is 0.06 mV/min after 5 h, indicating the stability and robustness of the chip. Moreover, the chip was applied to monitor pH changes in CaSki cells metabolism, with pH shifting from 8.04 to 7.40 on average. This platform has the potential for continuous and parallel monitoring of cell metabolism in single-cell culture arrays.
ASIC ISFET array pH monitoring ion imaging cell metabolism Journal of Semiconductors
2023, 44(2): 024101
1 安徽建筑大学环境与能源工程学院，安徽 合肥 230601
2 中国科学院合肥物质科学研究院安徽光学精密机械研究所，安徽 合肥 230031
3 环境污染控制与废弃物资源化利用安徽省重点实验室，安徽 合肥 230601
将三维荧光光谱技术与平行因子分析法相结合，实现了细胞培养基样本中色氨酸(TRY)、还原烟碱腺嘌呤二核苷酸(NADH)、黄素腺嘌呤二核苷酸(FAD)和维生素B6(VB6)含量的定性和定量分析。首先制备了25个细胞培养基样本，使用六因素五水平正交试验确定每个样本中四类荧光组分的混合方式和浓度梯度；采用荧光分光光度计扫描各样本的激发-发射矩阵，依次进行波长差异性校准、水拉曼和瑞利散射去除、内滤效应校正、光强标准化以及高杠杆点去除等预处理，然后采用水拉曼谱峰面积作为光强的当量单位，将荧光光强进行标准化；采用核一致矩阵确定模型的最佳组分数为4，最后使用平行因子分析法对三维荧光光谱数据集进行三线性分解，得到上述种荧光团的平均回收率分别为100.2%±15.5%、107.4%±37.1%、100.5%±6.4%、99.5%±6.5%，预测均方根误差(RMSEP)分别为0.124、43.312、1.601、0.639 μg/mL，相对平均偏差分别为13.216%、36.937%、6.112%、6.331%，检测限分别为0.013、52.628、0.003、0.012 μg/mL。TRY的发射光谱与NADH的激发光谱重合，且FAD的激发光谱与NADH的发射光谱重合，因此出现了荧光共振转移现象，该现象导致模型对NADH浓度的预测能力低于对其他三种荧光团的预测能力。模型在各荧光团的定性分析方面表现良好，均通过了S4C6T3的拆半分析校验。实验结果表明，三维荧光光谱结合平行因子分析法能够快速测定细胞培养基中多种代谢类荧光组分的含量，在细胞能量和物质代谢检测方面具有广阔的应用前景。
光谱学 三维荧光光谱 平行因子分析 代谢类荧光团 正交试验设计 内滤校正 拆半分析
1 西北农林科技大学 应用物理系,陕西 杨凌 712100
2 西安理工大学 应用物理系,西安 710048
为了从活体细胞角度阐述脉冲电场对作物萌发期抗旱性的影响及其机理,采用渗透势为－0.1 MPa的PEG-6000溶液形成干旱胁迫,研究了场强为100 kV/m、脉冲宽度为80 ms、脉冲频率为1 Hz的脉冲电场处理对干旱胁迫下萌发的玉米种子自发发光和延迟发光的影响.结果表明,在玉米萌发过程中,自发发光和延迟发光积分强度逐渐增长,而在干旱胁迫下降低,脉冲电场处理使干旱胁迫下萌发玉米种子的自发发光强度和延迟发光积分强度有所回升.自发发光强度和延迟发光积分强度的变化与玉米萌发抗旱指数和贮藏物质转运率的变化一致,表明脉冲电场处理提高了玉米萌发期的抗旱性,萌发玉米自发发光和延迟发光积分强度的变化分别从细胞呼吸代谢和细胞伤害的角度反映了脉冲电场对萌发玉米抗旱性的影响.研究结果揭示了脉冲电场对作物萌发期抗旱性的影响机理,可对开发基于脉冲电场的农业物理新技术提供参考.
超弱发光 脉冲电场 细胞代谢 玉米 抗旱性 无损检测 Ultraweak luminescence Pulsed electric field Cell metabolism Corn Drought resistance Non-destructive testing 光子学报
2015, 44(10): 1017002
为了揭示极低频脉冲电场对作物抗旱性的影响及其机理, 采用渗透势为-0.1 MPa的PEG-6000溶液模拟干旱条件, 研究脉冲频率为1 Hz 、场强为200 kV/m、脉宽为80 ms的极低频脉冲电场对干旱胁迫下玉米幼苗根系延迟光子辐射动力学参数的影响.结果表明, 在干旱胁迫下, 玉米幼苗根系细胞延迟光子辐射动力学参数的初始光子数I0、相干时间τ和衰减参数β均随着胁迫时间的延长呈现出波动性的下降趋势, 经过极低频脉冲电场处理的玉米幼苗根系细胞的初始光子数I0、相干时间τ和衰减参数β的变化趋势与之类似, 但是, 各动力学参数的数值均明显高于对照.延迟发光动力学参数的变化暗示极低频脉冲电场处理提高了渗透胁迫下玉米幼苗根系细胞的代谢水平和组织序性, 增强了细胞内功能分子之间的相互作用.延迟光子辐射的分析结果为在活体细胞层面阐明极低频脉冲电场对作物抗旱性的影响及其机理提供了参考.
延迟光子辐射 脉冲电场 渗透胁迫 根系细胞 细胞代谢 组织序性 抗旱性 Delayed biophoton emission Pulsed electric field Osmotic stress Root cell Cell metabolism Tissue order Drought resistance
为了解读植物种子萌发过程中超弱光子辐射信息的生物学意义, 采用呼吸抑制剂NaN3处理萌发玉米种子, 跟踪测量和分析了玉米种子萌发过程中超弱光子辐射中自发光子辐射和外界光诱导的延迟光子辐射的变化规律, 同时研究了萌发玉米种子鲜质量的变化.结果发现, NaN3同步抑制了萌发玉米自发光子辐射和鲜质量的增长, 造成萌发玉米延迟光子辐射的初始光子数和延迟光子辐射积分强度大幅度降低, 相干时间减小.机理分析表明, NaN3对呼吸代谢电子传递链的抑制造成的自由基反应减弱是萌发玉米自发光子辐射减小的原因, 自发光子辐射强度可以作为玉米萌发状态的信号, 延迟光子辐射动力学参数的大小可以表征萌发玉米呼吸代谢的强弱, 相干时间是种子细胞组织序性的量度, 通过对萌发种子超弱光子辐射的采集和分析可以实现对萌发种子细胞代谢和功能状态变化的灵敏和无损检测.
超弱光子辐射 种子萌发 呼吸抑制剂 细胞代谢 无损检测 玉米 Ultra-weak photon emission Seed germination Respiratory inhibitor Cell metabolism non-destructive testing Corn
