title: Compatibility of toluidine blue with laser microdissection and saturation labeling DIGE.
abstract: 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.
authors.affiliation: Wakefield Gastroenterology Research Institute, Wakefield Hospital, Wellington, New Zealand.
authors.forename: Chandra
authors.initials: C
authors.lastname: Kirana
authors.forename: Teresa
authors.initials: T
authors.lastname: Ward
authors.forename: T William
authors.initials: TW
authors.lastname: Jordan
authors.forename: Pisana
authors.initials: P
authors.lastname: Rawson
authors.forename: Janice
authors.initials: J
authors.lastname: Royds
authors.forename: Hong Jun
authors.initials: HJ
authors.lastname: Shi
authors.forename: Richard
authors.initials: R
authors.lastname: Stubbs
authors.forename: Kylie
authors.initials: K
authors.lastname: Hood
mesh_terms: D000704:Analysis of Variance; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004801:Eosine Yellowish-(YS) / Q000737:chemistry; D006416:Hematoxylin / Q000737:chemistry; D006651:Histocytochemistry; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D040901:Proteomics; D015203:Reproducibility of Results; D013194:Staining and Labeling; D014048:Tolonium Chloride / Q000737:chemistry*
chemical_list: D011506:Proteins; D014048:Tolonium Chloride; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin