A simple fluorescence of staining technique for in situ soil microorganisms.

A simple, rapid straining technique using the magnesium salt of 1-anilino-8-naphthalene sulfonic acid is described. Treatment of soil with an aqueous, membrane-filtered solution (3.5 mg/ml) of the salt causes the soil microorganisms to fluoresce when examined with light from a mercury arc light source.     

Techniques for staining fungal nuclei and appendages

PUNITHALINGAM, E., 1989. Techniques for staining fungal nuclei and appendages. The use of HCl-Giemsa stain for staining nuclei and a modified Leifson's flagellum staining technique for staining appendages in Coelomycetes has produced useful information which could further our knowledge of fungi and help to reappraise earlier concepts.     

Fluorescent feulgen staining of fungal nuclei

A rapid and alternative method for staining nuclei and chromosomes in fungal cells is reported. The method involves use of fluorescent dyes, specifically, auramin-O and acrinol as Schiff reagents. Micrographs depicting the distribution and division of nuclei in cells of four fungi are presented. These micrographs illustrate the potential of fluorescence microscopy for karyological investigations in fungi.     

A rapid whole-mount staining procedure for nuclei of mammalian embryos

A rapid staining procedure for examining nuclei of mammalian embryos is described. Embryos are placed on a glass slide, counterstained with trypan blue, stained with Hoechst 33342, and embedded between the slide and coverslip in Permount. The nuclei of the embryos fluoresce brightly when examined by fluorescence microscopy immediately after staining or after extended storage. The technique has proved to be an effective tool for studying the development of cow, hamster, mouse, pig, rabbit, and sheep embryos.     

A simple fluorescence staining technique for the differentiation of human tissue transplanted into nude mice.

Human and mouse nuclei can be distinguished by differences in the constitutive heterochromatin when stained with quinacrine dihydrochloride. With the staining method described, mouse heterochromatin during interphase appears as brilliant fluorescent chromocenters. By replacing the commonly used aqueous buffer mounting medium with a xylene-diluted synthetic resin, the haziness of the nuclear fluorescence is eliminated thus allowing identification of the heterochromatin pattern in histological preparations. A requirement for the definite identification of cells of human or murine origin in the nude mouse is the knowledge that the heterochromatin arrangements changes according to the stage of differentiation of the cell of the position of a particular nucleus within the cell cycle.     

A simple method for staining nuclei of mature and germinated maize pollen

A sugar acetocarmine staining technique has been developed for staining the sperm and vegetative nucleus of mature and germinated maize pollen grains. This procedure is simple, stable and highly repeatable. The physiological properties of the mature maize pollen grains are first adjusted by using an in vitro germinating culture solution. This solution is 15% sucrose and contains 360 ppm calcium chloride dihydrate, and 120 ppm boric acid. One part fresh pollen grains is uniformly mixed with nine parts of the solution and left at room temperature for at least 5 hr. One part of this solution is then mixed with two parts of regular acetocarmine stain and left overnight. The color of this mixture is pinkish red or raspberry. The sugar in the mixture helps to increase color contrast between the pollen cytoplasm (light pink) and the nuclei (reddish purple), decreases the frequency of burst pollen, increases pollen expansion, stabilizes pollen figures and automatically seals the coverglass.     

A hybrid technique for staining meiotic nuclei of Basidiomycetes.

A hybrid nuclear staining procedure used on the basidiomycete Schizophyllum commune produced squash preparations of sufficient density for effective brightfield photomicrography. Procedure: Tissure is fixed in Newcomer's solution, rehydrated in a graded ethanol series, rinsed, hydrolyzed in 1 N HC1 at 60 C, washed, mordanted in 4% iron alum, rinsed again, and stained with aceto-iron-hematoxylin prior to mounting and squashing.     

A rapid, simple method for staining bacterial flagella.

A simple modification of Gray's flagellar staining procedure is described. It can be used on air-dried smears or directly on wet mounts of motile bacteria. The stained bacterial flagella can be observed with phase-contrast or bright-field optics. No rigorous cleaning of slides, counterstains, or any washing procedures are required with the staining method, making it very suitable for routine examinations.     

A simple and rapid nuclear staining method for Rhizoctonia solani Kuhn

Abstract

We developed a modified staining technique using acridine orange to stain the nuclei of Rhizoctonia solani. Acridine orange solution was prepared in acetic acid buffer, pH 7.2. Staining for 15 min was critical for observing the nuclei. All of the isolates were found to be multinucleated. The nuclei appeared bright green with light orange background. This method is simple, rapid and reproducible.     

A new fluorescence technique for staining of mononuclear phagocytes.

A recently developed fluorochrome, 4',6-diamidino-2-phenylindole (DAPI), is used to stain mononuclear phagocytes of the mouse. After addition of heparin, these cells showed a bright yellow outer ring.     

A rapid and simple method for staining of the crystal protein ofBacillus thuringiensis

Summary A rapid and simple method of staining for the crystal protein (-endotoxin or parasporal body) ofBacillus thuringiensis has been developed. Changes in colonial morphology were observed when cells lost their ability to form crystal protein or both crystal protein and spore.     

A rapid, simple method for nuclei isolation from plant protoplasts

A rapid, simple method for nuclei isolation and purification from soybean (Glycine max L. Merr.) protoplasts is described. The isolated nuclei exhibited active amino acid incorporation and RNA synthesis, but DNA synthesis was not detectable. Analysis by CsCl density gradient centrifugation showed that DNA isolated from nuclei had a single band, while DNA isolated from protoplasts consisted of three bands comprised of nuclear DNA, mitochondrial DNA, and chloroplast DNA.     

A simple and rapid technique for assaying toxicants in aphids

A simple bioassay technique has been developed in order to study the effect of toxic compounds on aphids. The technique involves dipping of leaves of a particular aphid host in a solution of the desired toxic compound contained in a vacuum flask. The flask is then connected to a vacuum source in order to remove the air from the intercellular spaces of the palisade and the spongy parenchymas of the leaves. Breakage of the vacuum and return to atmospheric pressure forces the test solution into the now empty intercellular spaces which are thus filled with the solution. The treated leaves exhibited a typical water-soaked appearance. The infiltrated leaves were then offered to the aphids. The above treatment did not affect either the structure of the leaves or the development of the aphid colony on them. Aphids grown on leaves infiltrated with water lived longer in comparison with controls. In addition, insects grown on leaves infiltrated with 27.5, 13.7 and 6.9 p.p.m. of the systemic insecticide heptenophos died after 1.5, 3.0 and 5.0 h, respectively. The bioassay technique described in this report has certain advantages: it is simple, fast and inexpensive and is especially suitable for short-term studies involving the effects of such chemicals as toxins, insecticides and other toxicants on aphids. It may also be used to study the effects of growth factors, hormones and special nutrients on aphid growth and possibly on other sucking insects.     

PAS染色及真菌荧光染色在皮下真菌病诊断中的应用

皮下真菌病(subcutaneous mycoses)是指侵犯真皮、皮下组织和骨骼的真菌感染[1],部分可播及周边组织,如足菌肿等,有些则沿淋巴管扩散,如孢子丝菌病、着色芽生菌病,对人体造成较大危害。这类疾病病程缓慢,早期临床表现不典型,为疾病的早期诊断带来一定困难。真菌培养是其诊断的“金标准”,然而,真菌培养阳性率偏低,重要的是部分早期病例不容易考虑到真菌感染,多在组织病理提示后才考虑到深部真菌感染的可能性。