Histochemical Journal 

About: Histochemical Journal is an academic journal. The journal publishes majorly in the area(s): Staining & Acid phosphatase. It has an ISSN identifier of 0018-2214. Over the lifetime, 2705 publications have been published receiving 63835 citations.

Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections

Abstract: Sirius Red, a strong anionic dye, stains collagen by reacting, via its sulphonic acid groups, with basic groups present in the collagen molecule. The elongated dye molecules are attached to the collagen fibre in such a way that their long axes are parallel. This parallel relationship between dye and collagen results in an enhanced birefringency. Examination of tissue sections from 15 species of vertebrates suggests that staining with Sirius Red, when combined with enhancement of birefringency, may be considered specific for collagen. An improved and modified method of staining with Sirius Red is presented.

Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level.

Abstract: The argyrophilic proteins of the nucleolar organizer region (Ag-NOR proteins) were specifically localized at the optical level with a modified one-step silver technique performed at 20° C.

A new specific, sensitive and non-carcinogenic reagent for the demonstration of horseradish peroxidase

Abstract: 3,3'-Diaminobenzidine (DAB), introduced by Graham & Karnovsky (J. Histochem. Cytochem. 1966, 14, 291 & J. exp. Med. 1966, 124, 1123) is one of the most widely employed reagents in histochemistry. It has been very useful in demonstrating the sites to which the exogenous ultrastructural tracer horseradish peroxidase (HRP) is transported in vertebrate tissues. Over the past five years, DAB staining of transported HRP in neurons has developed into a powerful neuroanatomical tool for labelling the cells of origin of pathways in the central nervous system (La Vail, 1975, in: The use o f axonal transport for studies o f neuronal connectivity (eds. W. M. Cowan & M. Cuenod), pp. 217-248. Amsterdam: Elsevier.). Reports of the borderline carcinogenicity of DAB in rats (Griswold, Casey, Weisburger & Weisburger, Cancer Res. 1968, 28,924) may be responsible for the decreased availability of good quality DAB. Recent studies (Hanker, Anderson & Bloom, Science 1972, 175, 991; Hanker & Rabin, J. clin. Mierobiol. 1975, 2, 463) suggested that oxidative coupling reactions of aromatic amines in the presence of phenols might provide a suitable substitute for DAB. Some of these reactions yield deeply-coloured synthetic melanin-like compounds which are osmiophilic and sufficiently insoluble to be suitable end-products for histochemistry. In addition, the reaction must be sufficiently rapid to deposit the end-product at the sites of the plant hydroperoxidase alone. Such a reaction was realized when it was found that the peroxidation of p-phenylenediamine (PPD) was greatly accelerated by the presence of pyrocatechol (PC). The co-polymer formed as a result of this oxidative coupling reaction was osmiophilic and was bluer than oxidized DAB. It was insoluble and conformed well to biological ultrastructure. Neither PPD nor PC used individually gave satisfactory results and the best results were obtained when a mixture of 1 part (by weight) PPD to 2 parts PC was employed as the reagent. The administration of HRP and fixation of tissue for studies in mice, other than those involving axonal transport, were carried out according to Graham & Karnovsky (J. Histochem. Cytochem. 1966, 14, 291 &J. exp. Med. 1966, 124, 1123). For the demonstration of retrograde axonal transport in cats, rats, and rhesus monkeys, 30% HRP (Boehringer) in distilled water was injected into the sensorimotor cortex, or the ventrobasal complex of the thalamus (VB injection) or in the cerebellar cortex. The volume of HRP solution injected varied from 0.05 to 2/al. The animals were perfused with a mixture of 0.5% (para)formaldehyde and 2.5% glutaraldehyde in 0. t M phosphate buffer, pH 7.2. Tissue blocks (brain) were excised and immersed in 30% sucrose in 0.1 M phosphate buffer (pH 7.2) immediately after the perfusion. Cryostat or Vibratome* sections of mouse tissues were cut at 10-20/~m. Frozen sections of the rat, cat or monkey brains were cut at 40/2m.

Immunochemistry on ultrathin frozen sections

Abstract: Ultrathin frozen sections can be cut smoothly from many fixed and appropriately treated specimens. To use such sections for immunochemical localization of intracellular antigens, fixa ion conditions must be selected to optimize at least three variables, namely, preservation of ultrastructure, preservation of antigenicity and retention of accessibility of the antigen to the antibody. Furthermore, staining of the sections must be such that both the immunolabels and structures are clearly recognized. Our efforts to attain these goals are described in relation to their historical background. Although there are still problems to be solved and improvements to be made, we now consider that cryoultramicrotomy has reached the stage of being useful in studying many questions which will not be easily approached otherwise.

In situ detection of 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle tissue.

Abstract: Growing evidence suggests that intracellular vitamin D receptors are present in skeletal muscle tissue mediating vitamin D hormone response. The aim of the work reported here was to investigate the in situ expression of 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle tissue. Intraoperative periarticular muscle biopsies were taken from 20 female orthopaedic patients (17 middle-aged and elderly patients receiving total hip arthroplasty due to osteoarthritis of the hip or an osteoporotic hip fracture and 3 young patients who received back surgery). The immunohistological distribution of the vitamin D3 receptor was investigated using a monoclonal rat antibody to the receptor (Clone Nr. 9A7). The receptor-positive nuclei were quantified by counting 500 nuclei per biopsy. Strong intranuclear immunostaining of the vitamin D receptor was detected in human muscle cells. Biopsies of hip patients had significantly fewer receptor-positive nuclei compared to those of back surgery patients (Mann-Whitney U-test: p = 0.0025). VDR expression (number of antigen-positive nuclei) was significantly correlated with age (coefficient of correlation = 0.46; p = 0.005), but not with 25-hydroxyvitamin D or 1,25-dihydroxyvitamin D levels. The data clearly demonstrate presence of nuclear 1,25-dihydroxyvitamin D3 receptor in human skeletal muscle. To our knowledge this is the first in situ detection of the receptor in human skeletal muscle. The difference in the expression of the receptor between hip and spinal muscle biopsies might be explained by age or location. Further research is needed in order to evaluate whether vitamin D3 receptor expression in human skeletal muscle is age-dependent and varies between different muscles.