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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.

我们已经开发了微阵列化学传感器，它包括用于pH或氧气的光学传感器膜，以及用PDMS制备的微孔阵列，用于并行监测单细胞活性。在这项研究中，10克+ pH值和氧传感器点的安排，通过使用微接触印刷技术。制备的印模（点：10 μ m内径，间距：40 μ m）设置到微接触印刷机。用FITC和二氯三（1，10-菲咯啉）钌（II）分别作为pH和氧传感器试剂。将这些传感器试剂压印在类金刚石碳溅射载玻片上。采用激光共形高分辨率微阵列扫描仪测量每个点的荧光强度，将传感试剂渗透到透镜清洁纸巾中作为印台进行pH传感点的微接触印刷。平均斑点大小为10.0 ± 0.5 μm（n=48）。测量对pH 6-8的0.05M磷酸盐缓冲液的响应。100个点的平均荧光强度归一化的值在pH 7显示良好的相关性与pH值。对于氧传感器点的微接触印刷，传感器试剂滴到PDMS邮票，并通过使用气枪开发。平均斑点大小为9.9 ± 0.6 μm（n=25）。通过使用充有氧气或氮气的蒸馏水和5%亚硫酸钠溶液来评估传感器响应。99个点的平均荧光强度与氧浓度具有良好的相关性。最后，在氧传感器点印刷完成后，使用相同的PDMS印模将pH传感器点压印在氧传感器点之间。利用微阵列扫描仪测量了所制备的集成pH和氧传感器阵列的响应。每个传感器点都显示出特定的反应。