改良式免疫組織化學染色技術應用於福馬林液固定與石蠟包埋豬組織之研究

A Modified Immunohistochemical Staining Employed in Formalin-Fixed, Paraffin-Embedded Porcine Tissue Sections

劉振軒(Chen-Hsuan Liu) ； 王綉真(Hsiu-Cheng Wang) ； 蘇豐振(Feng-Chen Su) ； 傅千惠(Chien-Hui Fu)

《中華民國獸醫學會雜誌》 23卷6期 (1997/12) Pp. 523-530

https://doi.org/10.7009/JTCSVS.199712.0523

摘要

使用福馬林液固定與石蠟包埋之切片標本，以Avidin Biotinylated enzyme Complex （ABC）行免疫組織化學染色，常有費時過長且使組織抗原性不活化，因而降低染色品質之缺點，針對此缺失，乃進行改良式染色法研究以資改進。本研究係將豬大腦、小腦、肺、脾、肝、皮膚、心肌、淋巴結及腎等組織，經10%福馬林液固定與石蠟包埋切片後，配合MicroProbe（上標 TM）儀器及微波烤箱加熱處理，進行anti-human cytokeratin AE1/AE3, desmin, anti-swine vimentin, S-100 protein, anti-human glial fibrillary acidic protein（GFAP）, neuron specific enolase（NSE）及FIL-4（neurofilament 70 kd&200 kd）, factor VIII related antigen等八種抗體ABC免疫組織化學染色，並與傳統式不用MicroProbe（上標 TM）儀器與微波加熱處理的免疫組織化學染色作比較。結果顯示改良式免疫組織化學染色花費時間約需2-2.5小時，遠較傳統式花費時間過夜又5小時，不僅更具特效性，而且節省試劑與抗體之用量。在染色效果之比較，除了S-100 protein及factor VIII related antigen 改良式與傳統式並無顯著差異外，其餘anti-human cytokeratin AE1/AE3, desmin, anti-swine vimentin, GFAP, NSE及FIL-4（neurofilament 70 kd & 200 kd），改良式染色效果均較傳統式為佳。由上述實驗比較改良式與傳統式免疫組織化學染色結果，初步證實上述八種市售抗體均可適用上述兩種染色方法而應用於豬組織，使用改良式由於節省時間，染色效果穩定，所以是一種可行、方便並能爭取時效的免疫組織化學染色方法。

關鍵字

豬 ； 免疫組織化學染色 ； 微波加熱處理

並列摘要

Avidin Biotinylated enzyme Complex (ABC) immunohistochemistry employed in the formalin-fixed, paraffin-embedded porcine tissues is commonly seen. The disadvantages of formalin fixative and conventional staining method are antigen inactivation, time-consumption and reagent waste. For the purpose of improvement of staining effect and efficacy, we developed a modified staining method to compare with conventional staining method. In the modified method, 10% formalin-fixed porcine tissues including cerebrum, cerebellum, lung, liver, skin, myocardium, lymph nodes, and kidney were used, and ABC immunohistochemical staining of anti-human cytokeratin AE1/AE3, desmin, anti-swine vimentin, S-100 protein, anti-human glial fibrillary acidic protein (GFAP), neuron specific enolase (NSE), FIL-4 (neurofilament 70 kd & 200 kd), and factor VIII related antigen were performed by using microwave heating treatment and MicroProbe(superscript TM) The conventional method used neither microwave heating treatment nor MicroProbe(superscript TM). The results .showed that all above 8 market-available antibodies can be employed in porcine tissues by either modified or conventional method. However, the modified method took 2-2.5 hours compared with overnight of the conventional method. With respect to the staining effect, the modified method of anti-human cytokeratin AE1/AE3, desmin, anti-swine vimentin, GFAP, NSE, and FIL-4 (neurofilament 70 kd & 200 kd) staining showed a stronger and brighter effect than those stained with the conventional method. In conclusion, the modified immunohistochemical method is a practical, convenient, and fast technique to obtain reliable results.

並列關鍵字

Porcine ； lmmunohistochemistry ； Microwave heating ； M4icroProbe (superscript TM)