A fluorescence method for the visualization of inflammatory cells in intestinal mucosa.
Treatment of paraffin or cryostat sections from rat, rabbit and man, with fluorescein isothiocyanate (FITC) in buffer (1 microgram/l), resulted in visualization of highly fluorescent leukocytes with a granular staining in light microscopy. FITC staining of paraffin-embedded colonic tissue was compared with three other staining methods, i.e., (1) FITC-conjugated anti-neutrophil antibodies, (2) endogenous peroxidase with 3,3-diaminobenzidine, and (3) FITC-conjugated Lotus tetragonolobus lectin. Good correlations between FITC staining and the three other methods were found (r:0.97, 0.98 and 0.98; P less than 0.001). Inhibition of receptors for the FITC conjugates prior staining, by preincubation with unconjugated anti-neutrophil antibodies or L-fucose, abolished the staining with antibodies and lectin. This study shows that staining with unlabeled FITC represents a quick and convenient method for the estimation of phagocytic cells (polymorphonuclear cells and macrophages) in histological specimens.
Department of Pharmacology, Pharmacia AB, Uppsala, Sweden.
L G
LG
Axelsson
S
S
Ahlstedt
D000818:Animals; D004751:Enteritis / Q000473:pathology; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005453:Fluorescence; D007249:Inflammation / Q000473:pathology*; D007413:Intestinal Mucosa / Q000473:pathology*; D009504:Neutrophils / Q000473:pathology; D010586:Phagocytes / Q000473:pathology*; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling; D013861:Thiocyanates
D005452:Fluoresceins; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate