本實驗開發了一種三維有機電晶體(OECT)，透過蔻(coronene , CR)奈米纖維沉積於聚二氧乙基噻吩:聚苯乙烯磺酸(PEDOT:PSS)當作主動層通道，再經由後續DNA探針之表面改質，製作出一個能檢測miRNA的生物感測晶片。由於miRNA可由癌細胞萃取，估計此晶片可對於癌症病人之液態病理切片中的miRNA進行定量分析，首先我們利用真空熱蒸鍍製程將CR小分子沉積形成奈米纖維陣列於有機電晶體的聚二氧乙基噻吩:聚苯乙烯磺酸 (PEDOT:PSS)薄膜上，並利用π-π interaction將芘丁酸(PBA)經溶液修飾在CR鍍膜表面，然後利用EDC/NHS反應使 PBA的羧基得以和經胺基修飾的單鏈DNA (single - stranded DNA)探針接枝在晶片上，透過DNA和miRNA含氮鹼基互補對的特性，可以將待測miRNA的訊號放大，進行電訊號檢測。關於OECT元件效能分析方面，此實驗進行Id - Vd及跨導率 (transconductance)之量測，並分析其開關比例(on-off ratio)。關於miRNA感測方面，我們導入不同濃度下的miRNA的電流對響應時間的變化、ssDNA探針對miRNA的專一性做了檢測。此晶片對miRNA-21和miRNA-155的感測效能評估上，不僅能提供高靈敏度之量化感測，其最低檢測極限可達到10 fM，另外，利用和ssDNA探針不同序列的miRNA進行電流檢測，透過電流差異的大小，顯示其還可保有感測之專一性。期許對未來癌症之液態病理感測有極大幫助。

We have developed a three-dimensional organic electrochemical transistor (OECT) for the fabrication of a biosensor capable of detecting miRNA. The OECT utilizes coronene (CR) nanofibers deposited on a poly (3,4-ethylenedioxythiophene) : poly (styrenesulfonate) (PEDOT : PSS) layer as the active channel. Subsequently, the surface of the device is modified with DNA probes for the detection of miRNA through a series of modifications. First, CR small molecules are deposited as nanofibers array on the PEDOT:PSS thin film through a vacuum thermal evaporation process. Then, pyrenebutyric acid (PBA) is introduced onto the CR-coated surface through solution modification, taking advantage of π-π interaction. Next, the carboxyl groups of PBA are conjugated with amine-modified single-stranded DNA probes through an EDC/NHS reaction, allowing for the immobilization of the DNA probes on the chip surface. By exploiting the complementary base pairing between DNA and miRNA, the signal of the target miRNA can be amplified, enabling electrical signal detection. To evaluate the performance of the OECT device, measurements of Id-Vd and transconductance are conducted, and the on-off ratio is analyzed. Regarding miRNA sensing, we investigate the response time of the current signal with different concentrations of miRNA and test the specificity of the single-stranded DNA probes towards miRNA. The sensing performance of the chip is assessed for miRNA-21 and miRNA-155, demonstrating high sensitivity for quantitative detection with a limit of detection as low as 10 fM. Furthermore, the chip exhibits specificity in detecting miRNA by measuring the difference in current response between different sequences of ssDNA probes. This biosensor holds great potential for liquid biopsy analysis in cancer patients.

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