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Compatibility of toluidine blue with laser microdissection and saturation labeling DIGE
Author(s) -
Kirana Chandra,
Ward Teresa,
Jordan T. William,
Rawson Pisana,
Royds Janice,
Shi Hong Jun,
Stubbs Richard,
Hood Kylie
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800197
Subject(s) - laser capture microdissection , microdissection , h&e stain , eosin , staining , pathology , chemistry , microbiology and biotechnology , biology , biochemistry , medicine , gene expression , gene
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.