Cationic colloidal gold--a probe for light- and electron-microscopic characterization of acidic glycoconjugates using poly-L-lysine gold complex.
Cationic colloidal gold (CCG) was used to characterize acidic glycoconjugates in semithin and ultrathin sections of rat large intestine and salivary glands embedded in hydrophilic Lowicryl K4M resin. It was prepared from poly-L-lysine and 10 nm colloidal gold solution. The staining of CCG in semithin sections was amplified after photochemical silver reaction using silver acetate as a silver ion donor and examined under bright-field and epi-illumination microscopy. CCG adjusted to various pH levels was tested on various rat tissues whose histochemical characteristics with regard to acidic glycoconjugates are well known. At pH 2.5 CCG labelled tissues containing sialylated and sulphated acidic glycoconjugates such as the apical cell surface, mucous cells in the distal and proximal colon, and acinar cells of the sublingual gland. In contrast, CCG at pH 1.0 labelled tissues containing sulphated acidic glycoconjugates such as mucous cells in the upper crypt of the proximal colon and mucous cells in the whole crypt of the distal colon. This specificity of CCG was verified by the alteration of CCG staining following several types of cytochemical pretreatment. These results were further confirmed by electron microscopy. CCG staining is thus a useful postembedding procedure for the characterization of acidic glycoconjugates at both the light- and electron-microscopic levels.
Kashio
N
N
Department of Anatomy, Faculty of Medicine, Kagoshima University, Japan.
Tsuyama
S
S
Ihida
K
K
Murata
F
F
D002412:Cations; D003106:Colon / Q000737:chemistry / Q000648:ultrastructure; D006001:Glycoconjugates / Q000032:analysis*; D006046:Gold; D006651:Histocytochemistry / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D011107:Polylysine; D016622:Silver Staining; D013363:Submandibular Gland / Q000737:chemistry / Q000648:ultrastructure
D002412:Cations; D006001:Glycoconjugates; D011107:Polylysine; D006046:Gold