- 期刊
改良氏免疫組織化學染色技術應用於福馬林固定或酒精固定後石蠟包埋豬組織之研究
A Modified Immunohistochemical Staining Method Employed in Formalin or Alcohol-Fixed, Paraffin-Embedded Porcine Tissue Sections
摘要
本研究係將正常豬大腦、肺、脾、肝、皮膚、心肌、淋巴結及腎臟等組織採集並分別固定於10% 中性福馬林與絕對酒精。經石蠟包埋切片後配合Microprobe^(™)儀器，兩組切片以微波烤箱加熱處理，另二組未經微波烤箱加熱處理。進行Anti-human cytokeratin AE1/AE3，抗人類α 平滑肌肌動蛋白(anti-human asmooth muscle actin; Actin), Anti-human glial fibrillary acidic protein (GFAP), FIL-4 (neurofilament 70 Kd & 200 Kd), Anti-swine vimentin, Desmin，第八對因子相關抗原(factor Ⅷ related antigen; FactorⅧ)，Neuron specific enolase (NSE), S-100 蛋白質(S-100)，癌胚抗原(carcinoembryonic antigen; CEA)，板素(Iaminin)，抗人類髓鞘基礎蛋白(anti-human myelin basic protein; MBP)，抗人類巨噬細胞(anti-human macrophage; CD68)， 抗人類白血球共同抗原(anti-human leukocyte common antigen; CD45)等14種抗體免疫組織化學染色，比較不同固定液與微波加熱處理是否影響染色結果。切片經二位資深病理獸醫師作“盲目分析”判讀，記錄分數給與0-5 分不等染色強弱區分。結果顯示以10% 中性福馬林固定並以微波加熱處理與以絕對酒精固定且以微波加熱處理二組在Actin, GFAP, FIL-4, Vimentin 及NSE 等染色效果較佳。Desmin，S-100 及MBP則以10% 中性福馬林固定經微波處理染色效果最好。Cytokeratin 及Factor Ⅷ染色使用絕對酒精固定且不經微波處理，染色效果最好。CEA，板素，CD68，CD45 染色在四組均無法成功。
並列摘要
Modified avid in biotinylated enzyme complex (ABC) immunostain using 14 commercial primary antibodies were conducted in porcine cerebrum, lung , spleen , liver, skin, myocardium, lymph node, and kidney, which were either fixed in formalin or alcohol. These primary antibodies were anti-human cytokeratin AE1 /AE3 (Cytokeratin), anti-human α-smooth muscle actin (Actin), anti-human glial fibrillary acidic protein (GFAP), FIL-4 (neurofilament 70 kd & 200 kd), anti-swine vimentin (Vimentin), desmin, anti-human factor Ⅷ related antigen (Factor Ⅷ), neuron specific enolase (NSE), S-100 protein ( S-100), carcinoembryonic antigen (CEA), anti-human laminin (Laminin), anti-human myelin basic protein (MBP), anti-human macrophage (CD6S), and anti-human leukocyte common antigen (CD45). The tissues obtained from an adult pig were fixed in 10%neural buffered formalin or absolute alcohol at least 24 hours and routinely processed, treated with microwave unmasking, and stained using a Microprobe^(TM). Two groups only fixed in formalin or alcohol without microwave heating treatment were run in parallel as controls. The staining intensity was graded in a blinded fashion by two senior pathologists and was scored from zero to five. The results showed that 10 out of 14 antibodies were successfully applied in porcine tissues, whereas were negative for CEA, laminin, CD68, and CD45 antibodies. Under microwave unmasking procedure, tissues fixed in formalin or alcohol demonstrated stronger cytoplasmic positivity with cytokeratin, actin, GFAP, FIL-4, vimentin, and NSE antibodies than those stained without microwave unmasking. In comparison with effects of different fixatives with/without microwave heating treatment, desmin, S-100, and MBP antibodies using in formalin-fixed tissues plus microwave unmasking showed strong positivity. Fact or Ⅷ and cytokeratin antibodies in alcohol-fixed tissues without microwave unmasking showed strong positivity. In summary, commercial 10 antibodies of Cytokeratin, Actin, GFAP, FIL-4, Vimentin, Desmin, Factor Ⅷ, NSE, S-100 and MBP commonly employed in humans are also available in pigs using modified ABC immunohistochemical techniques. Different fixatives with/without use of microwave unmasking procedure may influence the staining intensity dependent on types of antibody.