Compatibility of toluidine blue with laser microdissection and saturation labeling DIGE.
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.
Kirana
Chandra
C
Wakefield Gastroenterology Research Institute, Wakefield Hospital, Wellington, New Zealand.
Ward
Teresa
T
Jordan
T William
TW
Rawson
Pisana
P
Royds
Janice
J
Shi
Hong Jun
HJ
Stubbs
Richard
R
Hood
Kylie
K
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*
D011506:Proteins; D014048:Tolonium Chloride; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin