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, rapid method for demonstrating bacterial flagella

We developed a simple, rapid method for demonstrating flagellation of bacteria using the fluorescent protein stain NanoOrange (Molecular Probes, Eugene, Oreg.). The NanoOrange reagent binds to hydrophobic regions of proteins, which results in substantial enhancement of fluorescence. Unbound reagent is essentially nonfluorescent. NanoOrange fluorescently stained bacterial cell bodies, as well as flagella and other appendages, which could be directly observed by epifluorescence microscopy. Detection of flagella was further improved by using a charge-coupled device camera for image capture and processing. The reliability of the method was tested by using 37 pure cultures of marine bacteria. Detection of flagella on the isolates by NanoOrange staining was compared to detection by transmission electron microscopy (TEM). For 36 of 37 cultures, the two methods yielded the same results. In one case, flagella were detected by TEM but not by NanoOrange, although the difference may be attributable to differences between the culture preparations. NanoOrange staining is rapid (10 to 15 min) and does not require fixation or dehydration, so live samples can be stained. Since NanoOrange is a general protein stain and works directly in seawater, it may also prove to be useful for staining other proteinaceous material that is of interest to aquatic microbial ecologists.     

A rapid staining technique for the detection of the initiation of germination of bacterial spores

AIMS: We propose to apply the Wirtz-Conklin staining technique to evaluate spore germination. METHODS AND RESULTS: Spores at different stages of germination were stained with modified spore stain (Wirtz-Conklin) and evaluated for staining properties. Bacillus spores suspended in deionized water, which does not support germination, stained greenish-blue. Spores suspended in germination enhancers that did not form bacilli stained pink, indicating the initiation of germination. Spores suspended in culture media, which promotes bacterial outgrowth, formed bacilli and were also stained pink. CONCLUSIONS: Modified spore stain (Wirtz-Conklin) was found to be useful to detect the initiation of spore germination as early as 30 min following incubation in a germination environment. SIGNIFICANCE AND IMPACT OF THE STUDY: This simple staining procedure is useful in detecting the initiation of germination of bacterial spores.     

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 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 simple method for the rapid removal of Bacillus anthracis spores from DNA preparations

This study establishes a filtration method for the safe removal of Bacillus anthracis spores which may contaminate DNA preparations. Centrifugal filtration with 0.1-microm filter units can be used following extraction of DNA from B. anthracis spores to render samples safe without compromising the sensitivity of diagnostic real-time PCR assays for B. anthracis.     

A rapid and simple staining method, using Toluidine Blue, for analysing mitotic figures in tissue sections

Summary Mitotic index is a clinically important parameter in cancer pathology. We developed a staining method using Toluidine Blue to detect efficiently and rapidly mitotic figures in sections of formalin-fixed paraffin-embedded human and rat tisues. Sections were stained at acid pH with a 0.01% Toluidine Blue solution after removal of RNA with hydrochloric acid or ribonuclease. The optimal pH of the TB staining solution was found to be 4.5 for rat tissues and 3.5 for human tissues. This procedure stained mitotic figures much more intensely than other (extra)cellular structures. A quantitative estimate of the total number of nuclei in the field where mitotic figures were counted, was obtained in an adjacent section hydrolysed in 5 N hydrochloric acid and stained by the Feulgen reaction with a Schiff-type reagent containing 0.01% Toluidine Blue. This method specifically stained interphase and mitotic nuclei and the field cellularity could be quantified by image cytometry. When these procedures were performed on two consecutive serial sections, a mitotic index could be determined accurately by relating the count of mitotic figures to the number of tumour cells.     

A simple and rapid in situ staining method for granulocyte-macrophage colonies in agar culture

This report describes a very simple, quick and effective method for in situ staining of granulocyte-macrophage colonies in agar cultures and for classifying various colony types. The procedure takes only one minute to fix and a few minutes to stain; a few additional minutes are required for preparation of the permanent whole plate. In this process the Riu stain, a modified Romanowsky stain, is used. Besides the ease and rapidity of this procedure, the identification of colony types appears to be enhanced. Thus, the method seems to be very beneficial in routine observations of colony types.     

A new, rapid, microwave-stimulated method of staining melanocytic lesions

A new and sensitive method of staining melanocytic lesions is described. Tissue sections covered by a solution of colloidal silver nitrate are exposed to microwaves for 45 sec in a domestic oven to produce clean, crisp staining of melanocytes and melanoma cells, often showing long delicate dendritic cell processes. The staining technique does not stain other pigments or argyrophilic tissues and is shown to be more sensitive than the standard Masson-Fontana procedure.     

A simple and rapid technique for fluorescence staining of fungal nuclei

A technique for staining fungal nuclei using fluorescence stain Hoechst Dye 33258 in McIlvaine standard buffer of pH 7.26–7.44 is reported. It is a broad-spectrum fungal nuclear staining tool found to be effective onAgaricus bisporus, Alternaria helianthi, Fusarium oxysporum f. sp.lini, Penicillium binellum, Pythium ultimum, Rhizoctonia solani, andSaccharomyces cerevisiae. Conidial nuclei ofAlternaria helianthi, Fusarium oxysporum f. sp.lini, andPenicillium binellum also stained well.     

A rapid and simple method for identifying bacterial polar lipid components in wet biomass

There are marked differences between wet and freeze-dried cells with regard to the identification of polar lipid components. The determination of the polar lipid composition of freeze-dried cells is well established. However, several approaches to identifying polar lipid components in wet cells have met with limited success owing to the presence of non-polar compounds in the extracts, resulting in a lipid composition with a narrow scope. In this study, we surveyed the lipid profiles of the wet biomasses of three Gram-positive (Microbacterium lacticum, Rhodococcus koreensis, and Streptomyces longwoodensis) and two Gram-negative (Pseudomonas aeruginosa and Novosphingobium capsulatum) bacteria; the results were comparable in quality to those obtained using a standard freeze-dried approach. Moreover, our improved method ensures simple lipid extraction. Overall, the results of the analysis showed minor lipid profile differences between the two approaches with regard to quantity, and lipid identification was consistent in both methods for all species.     

A simple rapid method for the detection of rat urinary proteins by agarose electrophoresis and nigrosine staining

The method described is sufficiently sensitive to detect major changes in the protein excretion patterns of rat urine, and the short time required for technical procedures makes the method suitable for screening large numbers of rat urine samples. The patterns observed for normal adult male rats are similar to previously published data, and the method may also be used to identify pseudoproteinuria.     

A rapid method for staining large chylomicrons

In this report, we present a rapid method for producing high-quality micrographs suitable for determining the size distributions of particles in concentrated samples of postprandial chylomicrons and chylomicron remnants. The procedure consists of mixing particles with osmium tetroxide in water to stabilize the lipids of the particles. These fixed and positively stained particles are then negatively stained with phosphotungstate in the presence of dilute sucrose. This dual staining procedure prevents the fusion and clustering of chylomicrons during processing for electron microscopy and is effective with particles of different lipid compositions. In addition, this procedure is simple and rapid, adding only one mixing step and 5 min to the preparation time required for conventional negative stains.     

优化的细菌鞭毛染色技术

优化了实验教材上传统的银染液鞭毛染色方法,用单宁酸和FeCl3做媒染剂,增大单宁酸和FeCl3的质量浓度(并将其配制的溶液分别保存),然后用碱性染料沙黄水溶液[1]、齐氏石炭酸碱性复红染液[1]和稀释10倍吕氏碱性美蓝染液[1],分别对培养好的枯草芽胞杆菌进行染色,得到较粗、清晰的染色结果。