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[求助] Lipofectamine2000+ASODN(FITC标记)的细胞摄取

本人用Lipofectamine2000+ASODN(FITC标记)做细胞摄取实验,L2000(微升)/ASODN(微克)=2.5/1和5/1,DMEM稀释,150微升每孔,ASODN的终浓度为1μg/ml。作用5个小时后,用pbs洗三次,在荧光显微镜下观察,结果无荧光,或者只有很小的绿色斑点,请各位同仁帮忙找找原因。
下面是20倍镜下拍的图片,
还有一个问题:细胞内的荧光是以眼睛看到的为准,还是以仪器拍到的为准?曝光时间不同,拍到的荧光图片差别很大,曝光时间一般设为多少?
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  • 附件 1 : L2000+A.tif
  • 2013-07-17 11:05:20, 14.11 M
  • 附件 2 : L2000(2.5-1)F.tif
  • 2013-07-17 11:06:37, 14.11 M
  • 附件 3 : L2000(5-1)F.tif
  • 2013-07-17 11:07:33, 14.11 M

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