To Make an Adequate Smear from Exudative Effusion by Cytocentrifugation

做出高品質的漏出液細胞離心抹片之條件

The report of a body fluid smear provides useful information for differential diagnosis, and it relies entirely on the good quality of smear. Many factors affect the quality of the smear. In this study, we had an isolated plasma, the mononuclear cells and the polymorphonuclear cells from whole blood. Then, we applied phosphate-buffered saline (pH = 7.4) and plasma to simulate the characteristics of exudative effusion (total protein ≧ 3.0 g/dL). We applied different filter pads, revolution per minute (RPM), and cellular suspension volume to investigate an optimal setting to make a good cytocentrifugated smear. The data of absorption duration for different filter pads were analyzed by t-test. Based on the smaller standard error of mean (SEM), the filter pad of the Shandon filter card (Thermo) was applied in the later experiments. The data of cell sizes, optimal centrifugation settings, and optimal sample volume settings were evaluated by line charts with standard deviations (SD). We concluded that the cell count of 1250 cells /μL, 400 μL of cellular suspension, high-speed acceleration and 700 RPM could make a high-quality smear when Shandon cytospin 4 and Shandon filter cards (Ref. 5991022) were applied in the cytocentrifugation for an adequate smear.

體液的細胞抹片鏡檢結果可以提供臨床醫師重要的鑑別診斷資訊，然而，這必須基於高品質的鏡檢報告。有許多因素會影響細胞離心抹片的鏡檢結果，在本項研究中，吾人從全血中分離出血漿、單形核細胞與多形核細胞，然後使用磷酸鹽緩衝液與血漿配置並模擬漏出液的特色（總蛋白質≧3.0g /dL）。吾人應用不同的細胞離心檢體杯濾紙、離心機轉速與檢體量找出最佳化參數以做成高品質的細胞離心抹片。不同細胞離心檢體杯濾紙的離心檢體量吸收時間以t檢定分析，並基於較小的平均值標準誤差而選定Shandon細胞離心檢體濾紙用於後續的實驗。細胞離心後的細胞大小、離心轉速的最佳化與細胞離心檢體量最佳化的數據分析是以折線圖搭配標準差而判定。經實驗結果分析，當使用細胞離心機Shandon cytospin 4並搭配Shandon細胞離心檢體濾紙，吾人判定細胞懸浮液的細胞濃度為1250 cells /μL、400μL細胞懸浮液、加速度設定為高值與離心轉速為每分鐘700轉可以做出高品質的體液離心細胞抹片。