Mechanism of connective tissue techniques. I. The effect of dye concentration and staining time on anionic dye procedures.
Anionic dye connective tissue procedures were performed by staining for 5 min and 24 h with (a) 0.00018 M and 0.0018 M solutions of 28 dyes, and 0.018 M solutions of 21 dyes in saturated picric acid (SPA), and (b) 0.0018 M and 0.018 M solutions of 20 dyes in 1% (w/v) phosphomolybdic acid (PMA). The staining obtained with dyes in SPA was classified as selective (no cytoplasmic staining), moderately selective (traces of cytoplasmic staining) and non-selective (all other staining patterns). The staining of collagen and cytoplasm with dyes in PMA was separately classified on a scale of 1-5 (1 = no staining, 5 = maximum staining). The selectivity of the staining obtained with SPA with solutions of dyes at concentrations of 0.00018 M and 0.0018 M, and both staining times, was correlated (p < 0.001) with an empirical sulphonic acid constant (SAC) defined as the (number of dye sulphonic acid groups/dye molecular weight) x 10(3). Correlation with molecular weight was poor and was significant only when staining was performed with 0.00018 M dye solutions for 24 h. The dyes were divisible into three groups: group 1 (selectivity independent, or almost independent of staining time), group 2 (selective to moderately selective when staining was performed for 5 min), and group 3 (non-selective). The SAC of the group 1 dyes differed significantly from those of the group 2 and 3 dyes. Selectivity was essentially lost at dye concentrations of 0.018 M. The staining with acidic dyes (no amines or substituted amines) in PMA differed significantly (p < 0.001) from that obtained with amphoteric dyes (containing basic substituents).(ABSTRACT TRUNCATED AT 250 WORDS)
Reid
P E
PE
Department of Pathology, Faculty of Medicine, University of British Columbia, Vancouver, Canada.
Iagallo
M
M
Nehr
S
S
Jankunis
M
M
Morrow
L
L
Trueman
L
L
D000818:Animals; D003094:Collagen / Q000032:analysis*; D003238:Connective Tissue / Q000737:chemistry*; D005456:Fluorescent Dyes; D007421:Intestine, Small / Q000737:chemistry; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008970:Molecular Weight; D008982:Molybdenum; D010756:Phosphoric Acids; D010851:Picrates; D051381:Rats; D017207:Rats, Sprague-Dawley; D012867:Skin / Q000737:chemistry; D013194:Staining and Labeling
D005456:Fluorescent Dyes; D010756:Phosphoric Acids; D010851:Picrates; D008982:Molybdenum; D003094:Collagen; C005858:picric acid; C003125:phosphomolybdic acid