MultiVysion® PB:所需试剂

MultiVysion® PB:所需试剂

简短版方案不应替代试剂盒说明书。

 

探针预混合在杂交缓冲液中。  每个目标区域使用 3 µL。 

 

注意:建议的最长杂交时间为 8 小时,但 4 小时为最佳。更长的杂交时间可能导致核染色偏高,从而可能淹没探针信号。

 

需要但不提供通用用途试剂。 

multivysion-pb-reagents-required.gif

 

请参阅样本制备部分以了解制备用于 FISH 的人卵裂球细胞的实验方案。

 
 
 
 
查看结果

MultiVysion® 杂交的适当查看和分析高度依赖于使用适当的滤光片组和校准良好的荧光显微镜。您的实验室中可能没有某些 MultiVysion PB 结果查看或成像所必需的滤光片组。请联系 Vysis 技术服务部门以了解有关滤光片组规格的更多信息。所有滤光片组均可通过 Vysis, Inc. 获得。 

 

建议使用以下最低配置滤光片组来查看 MultiVysion PB 结果:

 

1) 蓝色/青色双带通滤光片组

 

2) 绿色/红色双带通滤光片组

 

3) 金色(黄色)单带通滤光片组

 

4) 青色 v2 单带通滤光片组(可选,当难以区分蓝色和青色探针信号时用于辨别蓝色/青色双带通滤光片组中所观察到的信号)

 

请按以下方式查看杂交结果:

  1. 使用先前记录的坐标,在相位对比、低放大倍率 (100X) 下找到目标细胞。
  2. 一旦找到该细胞,在不移动显微镜载物台的情况下切换至使用青色单带通滤光片组的荧光灯,并在低放大倍率 (100X) 下重新定位细胞。当在荧光下重新找到该细胞后,切换至高倍率放大 (1000X) 进行分析。请根据将用于分析的滤光片组选择按照顺序 A 或 B 进行查看。 

A. 如果使用青色/蓝色双带通滤光片组分析 SpectrumAquaTM TM 和 SpectrumBlueTM TM 标记的探针,则请按以下步骤进行定位和分析: 

    • 使用青色/蓝色双带通滤光片组定位该细胞。
    • 按以下顺序使用滤光片分析其余每种探针:

1) 青色/蓝色双带通

 

2) 绿色/红色双带通

 

3) 金色(黄色)单带通

 

注意:建议您按照此顺序对样本进行查看或成像,以尽量减少光褪色的程度。按照此顺序,较敏感的荧光团将首先暴露于光下。

 

B. 如果使用单带通滤光片组进行分析,则请按以下步骤进行定位和分析:

    • 使用青色/蓝色单带通滤光片组定位该细胞。
    • 按以下顺序使用滤光片分析其余每种探针:

1) 蓝色单带通

 

2) 红色单带通

 

3) 金色(黄色)单带通

 

4) 绿色单带通 

 

5) 青色单带通

 
 
 
 

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