Development of cell chips equipped with micro-biosensing functions

我们已经开发了微阵列化学传感器，由pH或氧的光学传感器膜和PDMS制备的微孔阵列组成，用于并行监测单细胞活性。本研究采用微接触印刷技术布置了10个gm+ pH和氧传感器点。从SU-8制备的模具中复制PDMS印章。准备好的邮票（点：10um id，间距：40pm）被设置在微接触打印机上。FITC和二氯三（1,10-菲罗啉）钌（II）分别作为pH和氧传感器试剂。这些传感器试剂被印在钻石状的碳溅射玻片上。利用激光共角高分辨率微阵列扫描仪测量每个光斑的荧光强度。对于pH传感器斑点的微接触印刷，将传感器试剂渗透到透镜清洁组织中，并将其用作墨垫。平均光斑尺寸为10.0+0.5 μm （n=48）。测定对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与pH值归一化，与pH值具有良好的相关性。对于氧传感器点的微接触打印，将传感器试剂滴在PDMS印章上，并使用气枪显影。平均光斑尺寸为9.9+0.6 μm（n=25）。采用氧气或氮气的蒸馏水和5%亚硫酸钠溶液对传感器的响应进行了评价。99个点的平均荧光强度与普通蒸馏水的值归一化，与氧浓度有良好的相关性。最后，在氧传感器点打印完成后，在氧传感器点之间使用相同的PDMS图章进行pH传感器点的盖章。利用微阵列扫描仪测量了所制备的pH和氧传感器阵列的响应。每个传感器点都有特定的反应。

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.