開發流體動力學式微流道晶片應用於腹膜透析液中白血球計數

Abstract

腹膜透析是一種用來治療腎衰竭患者的透析療法。為了避免透析過程中可能會發生的腹膜發炎情形，監控透析液中白血球的比例及數目是非常重要的。傳統的檢測方式是使用流式細胞儀及自動細胞計數器對透析液中的白血球做定量，然而這些技術有以下的問題，像是(1)無法處理低濃度的樣本(<300 cells/μL)，(2)實驗儀器較為龐大且昂貴，這些問題限制了其使用性。為解決以上問題，本論文開發一利用流體動力學原理之微流道生醫晶片，此微流道系統操作簡單，製作成本低、製程簡單，可同時進行細胞、免疫微珠的抓取及分類。為了讓此微流道平台更加符合定點照護的需求，未來將與智慧型手機進行整合，並搭配軟體或是APP進行自動化影像處理、分析及資料傳輸。本論文主要分兩大部分，第一部分為免疫微珠抓取中性球之效率及能力測試，第二部分則為微流道分選並抓取白血球、免疫微珠的表現測試，未來則希望以Matlab軟體對白血球數目及微珠抓取細胞結果進行自動化定量。此微流道平台將能於定點照護的環境下偵測不同感染階段的腹膜炎，助醫生進行即時且正確的治療。

Peritoneal dialysis is a treatment for patients who suffer from severe chronic kidney disease. To prevent any infection during the treatment, it is important to monitor the population of white blood cell in peritoneal dialysis. At present, fluorescence-based flow cytometry and the automated hemocytometer are two prevailing methods to quantify the population of white blood cell. However, these techniques usually exist several limitations, such as (1) cannot deal with low white blood cell level (<300 cells/μL), (2) laborious assay preparation and manipulation steps. To address the above problems, we develop a microfluidic device with hydrodynamic trap arrays to capture white blood cells. The microfluidic microtrap array with multiple dimensions can trap general white blood cells and specific white blood cell subpopulation conjugated with 30μm polystyrene beads. This microfluidic platform enables simultaneously cell trapping, selection and can perform simple and real-time cell counting without complicated sample processing steps and equipment. To make the system more user-friendly and be suitable for point-of-care (POC) settings, we plan to make this microfluidic device compatible to existing smartphones and APP, which can perform image processing, analysis and data transmission. We believe this microfluidic platform for surveillance of white blood cell level will hold significant promise to provide the detailed infection status of patient for doctors to perform timely treatment.