對于一些特別堅硬或韌性比較強的樣品,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器中機器處理幾十秒鐘。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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