Improved Staining Method Research Articles

Flat, adherent, well-contrasted semithin plastic sections for light microscopy.

Semithin (0.5-2.0 μm) sections of plastic embedded specimens have long been used for identifying and orienting structures destined for electron microscopic observation. Improved staining methods and the development of more versatile plastics have increased the use of semithin plastic sections for histochemical and autoradiographic studies. The principal advantage of plastic over paraffin sections is the possibility of increased resolution. This advantage is often compromised, however, by problems arising during processing and staining. Wrinkles are common in sections containing tissues of different consistencies or when the hardness of the tissue does not match that of the surrounding plastic (Millonig 1980). Unfortunately, many of the methods designed to eliminate wrinkles (e.g., Alsop 1974, Sommer et al. 1979) require prolonged staining or repeated handling of the sections. Section adhesion problems usually arise during staining, particularly if the protocol requires alkaline or oxidizing reagents. Adhe...

Regional chromosome mapping of human collagen genes alpha 2(I) and alpha 1(I) (COLIA2 and COLIA1).

For the assignment of the genes for the pro-alpha 2(I) (COLIA2) and the pro-alpha 1(I) (COLIA1) collagens, cDNA and genomic DNA probes were used in in situ hybridization experiments on human prometaphase chromosomes. An improved staining method is reported for the simultaneous identification of chromosomes and the autoradiographic grains after the hybridization procedures. With this procedure more cells with higher resolution could be used for the assignment of genes by in situ hybridization. Statistical analysis of the grains located on respectively 660 and 302 metaphases using pro-alpha 2(I) and pro alpha 1(I) DNA probes, confirmed the assignment of these genes to human chromosomes 7 and 17. Analysis of the grain distribution on prometaphase chromosomes showed that the location of the pro-alpha 2(I) collagen gene is in the region 7q21.3-22.1. The location of the pro-alpha 1(I) collagen gene was found to be in band 17q21.31-2205.

A further study on the polymorphism of placental alkaline phosphatase in Japanese.

Polymorphism of placental alkaline phosphatase (PL) was investigated in 120 Japanese placentae by means of starch gel electrophoresis and an improved staining method for the isoenzymes using naphthol AS-MX phosphate as substrate instead of beta-naphthyl phosphate. The present method gave more satisfactory results for the detection of the PL bands. Besides five of the six common phenotypes three different uncommon variants were identified. The gene frequencies of PL types were estimated at PL1 = 0.729, PL2 = 0.054, PL3 = 0.200 and PLv = 0.017, which were very similar to those reported in other districts of Japan.

Quantitation and interaction of glycosaminoglycans with Alcian blue in dimethyl sulfoxide solutions

An improved method is described for the quantitation of glycosaminoglycans separatedon cellulose acetate, stained with Alcian blue, and dissolved in a dimethyl sulfoxide solution. Standard curves are shown for all eight glycosaminoglycans. It is shown that absorption at the Alcian blue orthochromatic E max is depressed under conditions which favor formation of dye-glycosaminoglycan complexes. The interaction between Alcian blue and the eight glycosaminoglycans was studied in dimethyl sulfoxide solutions of varying composition. It was shown that the extent of complex formation depends both on the glycosaminoglycan and the composition of the dimethyl sulfoxide solution. A dimethyl sulfoxide solution which contains 0.094 m H 2SO 4 is described which maximizes dye-glycosaminoglycan dissociation and thus the absorbance. Also, an improved staining method is described which improves dye uptake by the glycosaminoglycans and consequently increases the sensitivity of glycosaminoglycan quantitation.

C3, GBG, Orosomucoid and Haptoglobin Polymorphisms Improved Staining Methods

C3, GBG, Orosomucoid and Haptoglobin Polymorphisms Improved Staining Methods

C3, GBG, orosomucoid and haptoglobin polymorphisms. Improved staining methods.

C3, GBG, and orosomucoid polymorphisms were electrophoresed in a high-voltage agarose method which permitted the typing of 15-20 samples. The increased sensitivity of the dye Coommassie blue was used to stain the protein, and yielded higher resolution than amido black. The typing of haptoglobin samples, was facilitated by devising a method which utilizes the peroxidase activity of the haptoglobin-hemoglobin complex with 90-tolidine and hydrogen peroxide as substrates and 4-chloro-1-naphthol as coupler.

Studies on the microtubules in heliozoa. V. Factors controlling the organization of microtubules in the Axonemal pattern in Echinosphaerium (Actinosphaerium) nucleofilum.

On the assumption that the double-coiled pattern of microtubules in the axoneme of Echinosphaerium might be due to links of two sizes between adjacent microtubules, we disassembled microtubules with low temperature and then carefully analyzed the patterns of microtubules that formed upon the addition of heat (22 degrees C) or heat and D(2)O. Although most of the initial clusters of microtubules that formed could not be interpreted as part of an axoneme, the spacings between these microtubules were the same as that in the axoneme, 70 and 300 A. By model building we were able to show that all clusters that form, including stages in the formation of the axoneme and its 12-fold symmetry, could be explained by links of two sizes (70 and 300 A) and the substructure of the microtubule. We could demonstrate these links with improved staining methods. We suggest that nonaxonemal assemblies of microtubules may be eliminated by the natural selection of the most energetically stable configuration of microtubules, all others undergoing disassembly under equilibrium conditions. Model building further supports this suggestion since the model axoneme possesses more links per tubule than any other cluster found.

HISTOLOGICAL OBSERVATIONS ON THE RELATION OF INSULIN TO THE DEPOSITION OF GLYCOGEN IN ADIPOSE TISSUE1

The adipose tissue of animals in normal nutritional balance contains no histologically demonstrable glycogen. However, Gierke 1906 found that glycogen is deposited in the fat cells of guinea-pigs refed after a period of fasting. The occurrence of glycogen in the fat of other animal species under similar experimental conditions has been confirmed by several European investigators (Arndt, 1926, Wertheimer 1928, Richter 1931, Eger 1938, 1942). Among these Arndt, using dogs, also reported a transient accumulation of glycogen in the fat cells after insulin administration. These old observations take on new interest in the light of recent advances in our knowledge of the biochemistry of adipose tissue (Hook and Barron 1941, Mirski 1942), and the relation of insulin to lipogenesis (Drury 1940, Stetten and Klein 1946). We have therefore undertaken to investigate, with an improved staining method, the deposition of glycogen in the brown and white adipose tissues of normal and diabetic rats, rats given insulin, and...

An Improved Staining Method for Demonstrating Bacterial Capsules, with Particular Reference to Pasteurella

An Improved Staining Method for Demonstrating Bacterial Capsules, with Particular Reference to <i>Pasteurella</i>

The Kurloff-body, a spurious parasite

Twenty years or more have passed since Kurloff first observed certain vacuoles or cell enclosures in the protoplasm of the large mononuclear leucocytes of the guinea-pig while working in Ehrlich's laboratory. Since then improved staining methods and discoveries in the field of protozoology have made it possible to take new view-points in regard to these cell enclosures which are called Kurloff's-bodies. Therefore it is not with surprise that we find Ferrata, Patella, Goldhorn, Schilling and others believing these bodies to be protozoan in nature. Patella however is the only one who claims to have observed a flagellate develop from a Kurloff-body and of course this observation would put an end to all dispute if it could be verified.