對于一些特別堅硬或韌性比較強(qiáng)的樣品,如:皮膚、硬骨骼、軟骨、血管、結(jié)締組織、種子等,使用普通機(jī)械研磨方法很難即保證研磨破碎效果又保證樣品一直處于低溫狀態(tài)。傳統(tǒng)的手工研磨方法雖然能二者兼顧,但是當(dāng)樣品數(shù)量比較多時,手動研磨非常的費時費力,而且經(jīng)常存在浪費樣本、樣本間交叉污染的可能,影響后續(xù)實驗。如果提取RNA、酶等對溫度比較敏感的物質(zhì),使用液氮進(jìn)行預(yù)處理,保證樣品在破碎過程中一直處于低溫是非常重要的,上海凈信已經(jīng)成功解決這一問題。
使用上海凈信Tissuelyser全自動組織快速研磨儀進(jìn)行組織細(xì)胞的破碎時,將樣品用液氮進(jìn)行預(yù)冷凍,這樣既能保證樣品的破碎效率又能保證樣品一直處于低溫狀態(tài)。以血管和植物葉片為例,具體操作方法如下:
血管:
取新鮮血管,用剪刀剪成若干小塊,然后放入離心管中,再加入凈信研磨珠,蓋上管蓋。將整個試管放入液氮中進(jìn)行冷凍幾分鐘或直到液氮不沸騰,然后放入機(jī)器中處理幾十秒鐘。取出樣品管可見樣品被破碎成乳白色粉末,然后加入提取緩沖液,可以進(jìn)行接下來的后續(xù)提取,加入緩沖液之前能保證樣品溫度一直為-20℃以下。
葉片:
取植物葉片,清潔葉片表面后放入離心管中,加入幾顆凈信研磨珠,蓋上管蓋。將整個試管和適配器放入到液氮中冷凍幾分鐘或直到液氮不沸騰,然后取出放入研磨機(jī)器中機(jī)器處理幾十秒鐘。取出樣品管可見葉片被破碎成綠色粉末,然后加入提取緩沖液,進(jìn)行接下來的后續(xù)提取即可,加入緩沖液之前能保證樣品溫度一直為-20℃以下。
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