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Development of cell chips equipped with micro-biosensing functions

开发具有微生物传感功能的细胞芯片

基本信息

批准号：
18560750
负责人：
SUZUKI Masayasu
金额：
$ 2.53万
依托单位：
University of Toyama
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18560750/
关键词：
Micro-nano device Measurement Engineering Bioinstruments Microarray Bio Engineering

项目摘要

We have already developed micro-arrayed chemical sensors which consist of optical sensor film for pH or oxygen, and micro well array prepared with PDMS, for parallel monitoring of single cell activity. In this study, 10 gm+ pH and oxygen sensor spots were arranged by using microcontact printing technology.PDMS stamp was replicated from a mold prepared with SU-8. The prepared stamp (spot: 10 um i.d., pitch: 40 pm) was set to the micro contact printer. FITC and dichlorotris(1,10-phenanthroline) ruthenium (II) were used as pH and oxygen sensor reagent respectively. These sensor reagents were stamped onto a diamond like carbon sputtered slide glass. Fluorescence intensity of each spot was measured by using a laser conforcal high resolution micro-array scanner.For microcontact printing of pH sensor spots, sensor reagent was infiltrated into a lens cleaning tissue and it was used as an ink pad. Averaged spot size was 10.0+0.5 μm (n=48). Response to 0.05M phosphate buffer of pH 6-8 were measured. Averaged fluorescence intensity of 100 spots normalized with the value at pH 7 showed good correlation with pH. For microcontact printing of oxygen sensor spots, sensor reagent was dropped onto a PDMS stamp, and was developed by using an air gun. Averaged spot size was 9.9+0.6 μm(n=25). Sensor response was evaluated by using distilled water aerated with oxygen or nitrogen, and 5% sodium sulfite solution. Averaged fluorescence intensity of 99 spots normalized with the value at normal distilled water showed good correlation with oxygen concentration.Finally, after the printing of oxygen sensor spots, pH sensor spots were stamped between oxygen sensor spots by using the same PDMS stamp. The response of the prepared integrated pH and oxygen sensor array was measured by using a micro array scanner. Each sensor spot showed specific responses.

我们已经开发了用于并行监测单细胞活性的微阵列化学传感器，它由用于pH或氧气的光学传感器薄膜和用PDMS制备的微孔阵列组成。在这项研究中，利用微接触打印技术排列了10克+pH和氧传感器斑点，并在用SU-8制备的模具上复制了PDMS印章。将准备好的印章(点：10微米直径，节距：40 PM)设置到微型接触式打印机上。以FITC和二氯(1，10-邻菲咯啉)Ru(II)分别作为pH和氧敏试剂。这些传感器试剂被压印在一块钻石上，就像碳溅射的玻片玻璃一样。使用激光强制高分辨率微阵列扫描仪测量每个斑点的荧光强度，将传感器试剂渗透到镜片清洁组织中作为墨垫进行微接触打印pH传感器斑点。平均光斑大小为10.0±0.5μm(n=48)。测定了其对pH 6~8的0.05M磷酸盐缓冲液的响应。100个斑点的平均荧光强度与pH值有很好的相关性。对于氧传感器斑点的微接触打印，将传感器试剂滴在PDMS印章上，并使用气枪显影。平均光斑大小为9.9±0.6μm(n=25)。传感器的响应通过使用充氧或充氮的蒸馏水和5%亚硫酸钠溶液进行评估。99个点的平均荧光强度与正常蒸馏水中的氧浓度有很好的相关性，最后，氧传感器点打印后，在氧传感器点之间使用相同的PDMS印章来标记pH传感器点。利用微阵列扫描仪对所制备的集成pH和氧传感器阵列的响应进行了测量。每个感应点都有特定的反应。