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組織免疫染色

組織免疫染色. 研究部 黃欣儀. 組織製備方法. Paraffin embedded method Frozen section method Vibratome section method. Perfusion. Saline. PFA. Paraffin embedded method. 石蠟包埋法是將樣本經由固定、脫水、澄清、浸蠟後包埋在石蠟中，是製作光學顯微鏡切片的常規方法。它有許多優點，操作簡易便於掌握，可進行連續切片，包埋後的組織可以長期保存。 包 埋過程中須使用二甲苯，並且石蠟浸潤過程中需加熱處理。. Frozen section method.

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組織免疫染色

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  1. 組織免疫染色 研究部 黃欣儀

  2. 組織製備方法 • Paraffin embedded method • Frozen section method • Vibratomesection method

  3. Perfusion Saline PFA

  4. Paraffin embedded method • 石蠟包埋法是將樣本經由固定、脫水、澄清、浸蠟後包埋在石蠟中，是製作光學顯微鏡切片的常規方法。它有許多優點，操作簡易便於掌握，可進行連續切片，包埋後的組織可以長期保存。 • 包埋過程中須使用二甲苯，並且石蠟浸潤過程中需加熱處理。

  5. Frozen section method • 冷凍切片法是將樣本經由固定、抗凍脫水後包埋於冷凍切片劑中，是製作免疫切片的常規方法。優點是抗原保存性較佳，可切薄片，包埋後的組織可以長期保存。缺點是操作需技術，進行連續切片困難度較石蠟切片高。

  6. Cryoprotection • Protect biological tissue from freezing damage (damage due to ice formation). • Cryoprotectantsoperate simply by increasing the solute concentration in cells • In order to be biologically viable Cryoprotectantmust (1) easily penetrate cells, and (2) not be toxic to the cell • Common cryoprotectants: • Sucrose • DMSO (dimethyl sulfoxide) • Glycerol • Ethylene glycol • Propylene glycol

  7. Vibratomesection method • 振動切片法可針對新鮮或經過固定處理的樣本進行切片，樣本無需冰凍，是製作免疫切片的方法之一。優點是無冰晶形成，也可避免脫水、澄清及包埋對抗原的傷害，因此抗原的保存性最佳。缺點是樣本無法長期保存，切片染色需以漂浮法進行，而且無法進行薄切。

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