組織胺是引起過敏反應的主要物質之一，儲存於肥大細胞與嗜鹼性白血球的微粒中，臨床症狀包括氣喘、過敏性鼻炎、蕁麻疹，甚至休克等。在臨床上多以過敏原與特異性抗體進行體內皮膚試驗和體外血清過敏抗體進行診斷；然而，檢測過敏原與抗體僅能代表該個體對特定過敏原具有臨床反應，無法判定過敏反應發生之原因與其嚴重程度。因此本研究期望透過電化學檢測技術，針對細胞受到刺激後胞吐所釋放出的組織胺進行即時性偵測，藉以觀察並評估過敏反應時細胞的生理狀態。
研究共分為兩個部分: (1)以懸浮型人類嗜鹼性白血球細胞株(KU-812)為細胞模型，於鈣離子載體(calcium ionophore, A23187)刺激下釋放組織胺，並透過組織胺與o-phthalaldehyde (OPA)形成之複合物具螢光之特性，以冷光分析儀檢測組織胺釋放量。(2)開發電化學式微流體細胞檢測晶片，利用流力及自然沉降的力量將懸浮型KU-812細胞捕捉並安置於微孔電極(microwell electrode, MWE)中，以安培偵測法(+0.75 V vs. Ag/AgCl)進行組織胺分泌的電化學即時偵測。
從螢光檢測結果得知平均單一KU-812細胞內總組織胺含量約10~12 fmol，刺激30 min後分泌量約3~5 fmol。而微孔晶片實驗中，每次捕捉到1~3個細胞，刺激1~2 min後可觀測到一些不連續之突波電流(current spikes)，其響應電流約數個至數十個pA不等，響應時間約0.6~0.8 sec且持續約1 min。顯微鏡下觀察有突波個案者，其受測細胞均有胞吐的現象產生，將累積的突波電流之總電量進行計算，平均每個細胞約貢獻總電量2,000±200 pC，換算後約有6.7~26.8 fmol的組織胺被電極偵測到。本研究以電化學式微流體細胞檢測晶片，提供懸浮型細胞良好的檢測平台，未來可應用於模擬與評估不同類型細胞之生理狀態。

Histamine, which causes allergic inflammatory reactions, is one of the principal ingredients of mast cell and basophilic cell granules, and produces clinical syndromes that include asthma, rhinitis, urticaria, and even shock. The diagnosis of an antigen combined with a specific antibody is usually done through skin testing and blood testing in clinics. However, both tests are unable to judge the degree of damage when an allergy takes place, and only represent the clinical response for the individual. In this study, we used fluorometric assay to detect the coupling of histamine released from KU-812 with o-phthalaldehyde (OPA) and the resultant formation of a fluorescent product. For this, we developed an electrochemical cellular chip for continuous monitoring of histamine exocytosis from the suspension KU-812 cells. We found a low flow velocity area (LFVA) at the bottom of a microwell electrode (MWE), which could stabilize the trapped cell in a continuous flow, where the cells were accommodated and stabilized the MWE by the operation of hydraulic and gravitational forces. The released histamine was detected (+0.75 V vs. Ag/AgCl) by the MWE. Fluorescent result showed that each KU-812 cell contained 10~12 fmol histamine, and released 38% after being stimulated with calcium ionophore for 30 min, about 3~5 fmol. For amperometric detection of exocytosis at KU-812 cells, a noncontinuous current with randomly occurring spikes that had an average area of 2,000±200 pC and average histamine release of 6.7~26.8 fmol per cell was observed. Also, the exocytosis signals appeared after stimulated for 1~2 min, due to the exocytotic activity that can be stimulated by calcium ionophore by increasing the Ca2+ influx into cytoplasm. Both advantages of active (hydraulic) and passive (gravitational) traps were integrated in the proposed chip to avoid cells that were excited. The investigated cell could be stabilized within the MWE for electrochemical detection. Histamine was detected by the MWE, and oxidation current spikes still could be observed in A23187 stimulus, which can be used for living suspension cells detection. A spiked amperogram was recorded in activated KU-812 cells, and the spike current suggested the histamine release from exocytotic events. The proposed device demonstrated a capability for KU-812 cell analysis. The techniques presented here may be helpful for suspension cell array manipulation and electrochemical measurement for cell biological studies.

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