Quantitative amplification of genomic DNA from histological tissue sections after staining with nuclear dyes and laser capture microdissection
Quantitative amplification of genomic DNA from histological tissue sections after staining with nuclear dyes and laser capture microdissection
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DOI：
10.1016/s1525-1578(10)60645-9
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发表时间：
2001-02-01
影响因子：
4.1
通讯作者：
Watson, MA
中科院分区：
文献类型：
--
作者：
Ehrig, T;Abdulkadir, SA;Watson, MA
激光捕获显微解剖(LCM)允许选择性地从组织切片中采样组织。这项技术的一个先决条件是获得不会干扰样本遗传物质的下游分析的组织学染料。我们研究了四种组织学核染料(甲基绿、苏木素、甲苯胺蓝O、天青B)对TaqMan聚合酶链式反应扩增β-肌动蛋白的影响。从固定和冷冻的组织中制备的基因组DNA。将手工解剖取自组织切片的组织与激光共聚焦显微镜(LCM)采集的组织进行比较。正如以前报道的，在分析手工解剖的组织切片时，苏木素染色后的DNA聚合酶链式反应扩增效果不如其他染料染色后的DNA扩增结果。反之，用激光共聚焦显微镜检测时，苏木精染色后的DNA回收率与甲基绿染色后的相当。我们的结论是，从LCM样本组织中回收DNA不依赖于为突出核细节而选择的组织学染色。
Laser capture microdissection (LCM) allows the selective sampling of tissue from histological sections. A prerequisite for this technique is the availability of histological dyes that do not interfere with downstream analysis of the sampled genetic material. We have examined the effect of four histological nuclear dyes (methyl green, hematoxylin, toluidine blue O, azure B) on TaqMan polymerase chain reaction amplification of beta -actin. genomic DNA prepared from fixed and frozen tissue. Tissue sampled from the histological sections by manual dissection was compared with tissue sampled by LCM. As previously reported, when manually dissected tissue sections were analyzed, polymerase chain reaction amplification of DNA after hematoxylin staining was inferior to that after staining with the other dyes. In contrast, when tissue sampled by LCM was examined, DNA recovery after hematoxylin staining was equivalent to the recovery after methyl green staining. We conclude that DNA recovery from LCM-sampled tissue is independent of the histological stain chosen to highlight nuclear detail.