臨床角膜潰瘍病患檢測流程中，從取得病灶組織再送至病理室進
行感染微生物培養與鑑定常耗時數天至數周。拉曼光譜因具有微量生物樣本檢測與快速分析之特點，近年來於微生物檢測之應用逐漸受到矚目。雖然拉曼光譜可直接針對病灶檢體進行檢測分析，可大幅縮短檢測時間，然其獲得之光譜訊號因相當複雜，致使解析與結果的判斷不易。此研究以角膜潰瘍病患檢體分析後發現，患者淚液及病灶組織刮除後之眼表清洗樣本，常因細菌濃度過低而不易於拉曼光譜之檢測，故本研究藉由「Coffee ring effect」作用，透過控制拉曼基材上的樣本液滴乾燥過程，而達到使細菌濃縮與聚集效果。實驗結果發現，樣本於垂直放置之基材上並配合適當的乾燥時間，可具有較明顯的菌體聚集效果，若再配合光譜mapping 模式可於低濃度細菌樣本中獲得較佳的細菌指紋圖譜和檢測效果。

Traditionally, corneal ulcer is often examined by culture to determine the kind of infected microorganisms. However, it is time-consuming. Recently, there are many reports regarding Raman spectrum analysis to try to promote such a test, because of the rapid detection and analysis under the Raman detection. Although Raman spectra can detect real
sample directly and reduce the examination time. But it’s hard to identify the signal. In this study, we try to use such a method to analyze tears of corneal ulcer patient infected by bacteria. Owing to the rare bacterial
population in the tear sample that may be obtained from patient, a suitable pretreating method based on coffee ring phenomenon is needed. According to coffee ring effect, the latex samples simulated as tear samples with bacteria were dropped on the gold-coated glass slide, and
then we made them to be 90° standing for drying in the air. A lot of latex can be collected at the bottom of edge. From the results, we could compare the effects by the different angles of substrates and the different
periods of time to dry in air. It could achieve concentration and aggregation of the sample. Through improving the aggregation effect at low concentration of the sample, this method may also provide us to detect the Raman spectra of the real tear samples with microorganisms.

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