Kirana Chandra, Ward Teresa, Jordan T William, Rawson Pisana, Royds Janice, Shi Hong Jun, Stubbs Richard, Hood Kylie
Wakefield Gastroenterology Research Institute, Wakefield Hospital, Wellington, New Zealand.
Proteomics. 2009 Jan;9(2):485-90. doi: 10.1002/pmic.200800197.
Tissue fixation and staining protocols for laser microdissection are frequently not fully compatible with subsequent proteomic analysis. We compared the effect of three common histological stains (toluidine blue (TB), hemotoxylin, and hematoxylin and eosin (HE)) on tissue visualization, protein recovery, the saturation labeling reaction, and 2-D electrophoresis. TB provided the best visualization of colorectal tumor tissue during laser microdissection (LMD) and had a comparable effect on protein recovery and the saturation labeling reaction with hematoxylin, provided a modified 2-D clean-up protocol was used. Eosin inhibited both protein recovery and the saturation labeling reaction.
用于激光显微切割的组织固定和染色方案通常与后续的蛋白质组学分析不完全兼容。我们比较了三种常见组织学染色剂（甲苯胺蓝（TB）、苏木精和苏木精-伊红（HE））对组织可视化、蛋白质回收、饱和标记反应和二维电泳的影响。在激光显微切割（LMD）过程中，TB能提供结直肠肿瘤组织的最佳可视化效果，并且在使用改良的二维清理方案时，对蛋白质回收和饱和标记反应的影响与苏木精相当。伊红则抑制蛋白质回收和饱和标记反应。
