STUDIES ON THE PHYSICAL DEVELOPER FOR USE IN IMMUNOHISTOCHEMISTRY

• Published in: ACTA HISTOCHEMICA ET CYTOCHEMICA Vol. 23; no. 5; pp. 647 - 662
• Main Authors: TAKITA, KAORUKO, SHIMADA, TATSUO, KITAMURA, HIROKAZU, FUJII, SHIGETADA, NAKAMURA, MITSUO
• Format: Journal Article
• Language: English
• Published: Kyoto JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY 01.01.1990; Japan Society of Histochemistry and Cytochemistry; Japan Science and Technology Agency
• Subjects: Biological and medical sciences; Diverse techniques; Fundamental and applied biological sciences. Psychology; Molecular and cellular biology; Performance evaluation; Methodology; Rat; Rodentia; Peptide hormone; Tissue culture; Insulin; Neuropeptide; Citric acid; Dose activity relation; Vertebrata; Optical microscopy; Mammalia; Malic acid; Histochemistry; Coloration
• ISSN: 0044-5991; 1347-5800
• DOI: 10.1267/ahc.23.647

Four experiments were carried out to investigate physical developers containing bromohydroquinone. In experiment 1, the effect of citric acid concentrations in physical developers upon the intensity of immunohistochemical reactions was investigated, using a protein A-gold silver technique for detection of insulin. It was noted that the reactions were decreased in intensity with increasing concentrations (300, 400, 500 and 700mg/61ml) of citric acid involved. Addition of 250mg/61ml sodium citrate to the developer containing 700mg/61ml citric acid elevated pH to a value nearly comparable to that of the developer containing 300mg/61ml citric acid. Such a maneuver increased the intensity of the reaction, however, not to a level reached when the developer containing 300mg/61ml citric acid was used. Thus, it is considered that citric acid controls the developing capacity not only by modulating pH values but by complexing with silver. In experiment 2, the possibility was substantiated that the citric acid involved can be replaced with maleic acid. In experiments 1 and 2, we newly prescribed two refined physical developers (Developers 2 and 3). Developer 2 contains a larger amount (500mg/61ml) of citric acid as compared with Developer 1 which we have thus far employed primarily. In Developer 3, maleic acid is contained instead of citric acid. In experiment 3, these revised physical developers were used in protein A-gold silver or immunogold silver staining for detection of a series of antigenic substances, and proved to be excellent, in terms of the intensity, sensitivity and specificity of histochemical reaction. Further, experiment 4 in which Danscher's physical developer was tentatively used in immunostaining substantiated the superiority of the present revised developers to the one elaborated by Danscher (7).