108 results about "ANILINE BLUE" patented technology

The invention relates to a method for observing microscopic structures inside plant roots, which is capable of effectively solving the problem of utilizing a microscope to clearly observe the structures inside the plant roots. The method comprises the steps of digging out the plant roots, washing, shearing the plant roots into small sections, fixing the plant roots by methanol, and then embedding by a sepharose solution, cooling, condensing, trimming into small blocks, after freehand section, placing the small blocks into distilled water in a container, performing lactic acid clearing, covering, dyeing via berberine or fluorescein, bleaching by the water, performing poststaining via toluidine blue O or sarranine O, bleaching by the water, performing water seal, observing and photographing via an inverted fluorescence microscope, and obtaining a clear image. The method can be used for observing the microscopic structures inside the plant roots, thereby being effectively used for the research on classification, auxanology, ecology and physiology of medical plants. The method disclosed by the invention has the advantages of simple method, easiness in operation, low cost, low working capacity, high success rate, and good effect, thereby being capable of being used for the research on morphological anatomy of the roots. The method is simpler and more effective for the thinner root sections of the plants.

The invention belongs to the field of biotechnology, and discloses a laccase gene as well as an encoded protein and an application thereof. The open reading frame sequence of the laccase gene fmb-L103 is shown in SEQ ID NO. 1. The amino acid sequence of the protein encoded by the laccase gene fmb-L103 disclosed by the invention is shown in SEQ ID NO. 2. The laccase gene fmb-L103 disclosed by the invention is applied to decoloration for triphenlmethane dyes. The inventor clones from the selected dead bacillus vallismortis fmb-L103 with a spore laccase activity to obtain a novel prokaryotic laccase gene fmb-L103, and realizes the heterologous efficient expression of escherichia colli expression host bacteria. The recombinase fmb-L103 can be used for efficiently catalyzing the degradation of the triphenlmethane dyes of malachite green, brilliant green and aniline blue.

The invention provides a method for quickly identifying cross-compatibility of rose hybrida. The method is characterized by comprising the following steps of: collecting and storing pollen, performing castration, performing artificial pollination, cutting a sample to be identified, fixing the cut sample in FAA stationary liquid, transferring the cut sample to a 70 percent ethanol solution after 24 hours, taking out for washing, placing in an 8mol/L NaOH solution, performing water bath at the temperature of 55 to 60 DEGC for 15 to 20 minutes, and then standing at the temperature of 20 to 28 DEGC for 3 to 3.5 hours, soaking into water for at least 1 hour, transferring to a 0.1 percent dye solution of aniline blue, dyeing for 2 to 3 hours, observing stigma callose deposition, pollen germination and growth of pollen tubes by using a fluorescence microscope to identify the cross-compatibility of rose hybrida. By the method, the cross-compatibility of rose hybrida can be identified within 30 to 48 hours; and moreover, the method is simple and easy to operate and can effectively reduce the waste of breeding resources and improve breeding efficiency.

The invention discloses a screening method of a high-performance hydrolytic bacterial strain of waste Baijiu distiller grains. The screening method is characterized by comprising the following steps:step I, directly separating and culturing fungus from effective fungus that possibly hydrolyzes waste Baijiu distiller grains of relevant waste hydrolytic Baijiu distiller grains in a bacterial strainlibrary in a waste Baijiu distiller grains stacking environment; step II, performing the primary screening for the screened fungus successively by virtue of a sodium carboxymethylcellulose-Congo redculture medium, PDA-aniline blue culture medium, and a xylan-Congo red culture medium by adopting the single polymer which is difficult to hydrolyze in the waste Baijiu distiller grains as an exclusive carbon source; step III, culturing the bacterial strain in a waste distiller grain solution adopting the waste Baijiu distiller grain as an exclusive nutritional substance, and determining the capability of the bacterial strain for the waste distiller grains; step IV, performing the enzyme activity test for the composite bacterial strain in the waste Baijiu distiller grain solution, and screening the bacterial group with synergistic effect and high enzyme activity, thus obtaining the high-performance composite hydrolytic bacterial group; and step V, comparing enzyme activity and biomass hydrolytic capability of the high-performance composite bacterial strain with that of the disclosed mutant bacterial strain having the high-performance hydrolytic cellulose.

The invention discloses a method for determining the ion content of cationic polyacrylamide by utilizing spectrophotometry, which is characterized in that polyvinyl alcohol lemery is prepared into a standard solution, a colloidal titration method of titrating a cationic polyacrylamide dilute solution is adopted, and visible light spectrophotometry is adopted to judge the end point of the titration. The method comprises the concrete steps: 1. preparing the standard solution; 2. preparing the cationic polyacrylamide solution and diluting; 3. regulating the pH value to 1-3 with 1 percent of hydrochloric acid, and adding 100 microlitres of 0.5 percent of toluidine blue indicator; 4. titrating with the standard solution; 5. drawing a photometric titration curve S; 6. and doing a group of blankexperiments by referencing the steps of 1-5. The invention uses the photometric titration to determine the ion content of cationic polyacrylamide and observes the color change of the solution insteadof naked eyes, thereby reducing end point judgment errors and avoiding the determining failure caused by color reversion, and therefore, the goals of increasing the end point judgment precision and the determining result accuracy are achieved.

The invention discloses a method for detecting β-glucan based on the principle of fluorescence enhancement. The treated silicon wafer is used as the substrate, and titanium dioxide and graphene oxide hydrogel are deposited on the silicon wafer in combination with spin-coating technology to prepare titanium dioxide/ Graphene oxide hydrogel one-dimensional photonic crystal, and gold-plated on the photonic crystal by vacuum evaporation method, prepared gold-plated titanium dioxide/graphene oxide hydrogel one-dimensional photonic crystal, and used this photonic crystal as a fluorescence-enhanced Substrate, a method for detecting β-glucan by detecting the fluorescence intensity of the fluorescent complex product specifically combined with aniline blue. The present invention uses gold-plated titanium dioxide/graphene oxide hydrogel one-dimensional photonic crystals as the base, and combines the fluorescence enhancement effects of gold nanoparticles and photonic crystals to cause greater fluorescence enhancement, thereby improving the sensitivity of fluorescence detection. The method is simple and effective, the operation is convenient and the required time is short.

The invention relates to thermophilic aerophilic thiamine-decomposing bacillus, a microbial inoculum and application of the microbial inoculum. The bacillus can well grow at the temperature of 30-60 DEG C. In an aniline blue decoloration test, a decoloration ring with the diameter of 5cm can be generated within 24h. The microbial inoculum has the advantages of high lignin degradation rate, high temperature resistance, short growth period and the like when being applied to lignin degradation, and particularly can be used for strain resources of high-temperature straw compost.

The present invention is directed to a novel multi-spectral contrast agent-enhanced polarized light imaging technique that enables rapid imaging of large tissue fields. The imaging device includes a tunable monochromatic light source and a CCD camera. Linear polarizers are placed into both the incident and collected light pathways in order to limit the measurement volume to the superficial tissue layers. To enhance the tumor contrast in the image, aqueous solutions of toluidine blue or methylene blue are topically applied.

The invention discloses a catalytic reduction hemoglobin electrochemical sensor which can achieve fast and sensitive detection of hemoglobin in a relatively large concentration range; the electrochemical sensor is a three-electrode system composed of a working electrode, a reference electrode and a counter electrode; the working electrode is prepared by the method according to the following steps: putting a three-electrode system with a glassy carbon electrode as the working electrode into a sodium acetate-acetic acid buffer solution having the pH of 5.0 and containing graphene and toluidine blue, and in a potential range of -0.8 to 1.3 V, carrying out cyclic voltammetry scanning for 25 laps at the scanning speed of 100 mV/s, to prepare a graphene/polytoluidine blue modified electrode; and putting a three-electrode system with the prepared graphene/polytoluidine blue modified electrode as the working electrode into an aqueous solution containing chloroplatinic acid and sodium chloride, and applying a -0.7 to -0.8 V constant potential between the working electrode and the reference electrode with the application time of 60-240 s.

The invention discloses a method for degrading a triphenylmethane dye by utilizing recombinational lipoxygenase and application of the recombinational lipoxygenase rLOX-27853 in degrading the triphenylmethane dye. The amino acid sequence of the recombinational lipoxygenase rLOX-27853 is as shown in SEQ ID NO.2. The method for degrading the triphenylmethane dye by utilizing the recombinational lipoxygenase comprises the following steps: adding the recombinational lipoxygenase rLOX-27853 to a medium which contains sodium linoleate and the triphenylmethane dye to degrade the medium. The method disclosed by the invention obtains a novel prokaryotic lipoxygenase gene LOX-27853 by cloning from a pseudomonas aeruginosa ATCC27853 strain genome and realizes the heterogenetic high-efficiency expression of host bacteria in escherichia coli by utilizing a fermentation culture method, thereby efficiently catalyzing the degradation of the triphenylmethane dyes, namely aniline blue, malachite green and brilliant green.

The invention relates to a method for treating paddy rice tissues by using a prokaryotic expression product of a magnaporthe grisea effect protein gene and inducing generation of callose, belonging to the technical field of plant protection and biology. The method is an improvement on the prior art, comprises a prokaryotic expression technology which is the same as that of the conventional method, a protein purifying technology, the wavelength ranges of exciting light and emitted light of a used fluorescence microscope and an aniline blue dyeing method, and is characterized by comprising the following steps of: treating the root tissues of Nipponbare serving as a disease-resistant paddy rice variety with the prokaryotic expression product which is 1.0-10.0 mg/ml in concentration of the magnaporthe grisea effect protein gene is used for treating the root tissues for 12 hours; performing aniline blue dyeing; and directly observing the formation of callose under the fluorescence microscope. The method is easy, convenience, accurate and rapid; and a large amount of spontaneous fluorescence interference can be avoided effectively, and the situation of basic defense reaction of paddy rice can be obtained directly and qualitatively.

Methods of detecting progression from precancer to cancer are provided utilizing toluidine blue staining as well as detecting allelic variation at microsatellite loci. An allelic variation in one or more locus is indicative of a progression from precancer to cancer.

The invention discloses a method for identifying self-fruitful pear varieties by using pollination in vitro indoor, belonging to the field of plant breeding. The method comprises the following steps of: carrying out self-pollination by using large flower buds; immediately cutting styluses from the base parts of the styluses and the ovaries by using blades after carrying out the self-pollination; placing the styluses on a solid medium, culturing the styluses for 48 hours under the dark environment of 25DEG C, and taking out the styluses; dripping several drips of 0.1 percent aniline blue staining fluid on the base parts of the styluses; and observing the grow of pollen tubes from the base parts of the styluses under a microscope. If the pollen tubes grow from the base parts of the styluses, the variety can be fruitful by using the self-pollination; and if the pollen tubes do not grow from the base parts of the styluses, the variety cannot be fruitful by using the self-pollination. The method solves the problem of identifying the self-fruitful property in large quantity within one year, not only can identify all pear variety resources, but also can reference the identification of the self-fruitful property of other fruit trees and has wide application prospect.

The invention provides a non-toxic coloring agent used for detecting grass endophytic fungi. The non-toxic coloring agent comprises aniline blue, lactic acid and solvent. The non-toxic coloring agent is free from phenol harmful to human beings and environment, coloring effect is improved by simply heating slides on alcohol lamp flame during coloring, and distinguish between endogenous fungal hyphae and cell walls is facilitated.

The invention discloses a composite dyeing agent and an application thereof. The composite dyeing agent is mixed liquid consisting of an acetic acid solution, a toluidine blue solution and normal saline, wherein in the mixed liquid, the mass concentration of acetic acid is 1.5%-2%, and the mass concentration of toluidine blue is 0.2%-0.4%. The composite dyeing agent cooperates with a gastroenterological endoscope, so that the focus of a disease, which is difficult to distinguish can be convenient to find, and the composite dyeing agent is particularly used for esophagitis of dogs, and can improve the diagnosis rate and the sensibility of the esophagitis of the dogs.

The invention relates to a method for evaluating the development situation of cuticles of mature apple fruits. The method is characterized in that the apple fruits undergo aniline blue dyeing under a 0.04-0.1MPa vacuum condition, and the water absorbing capacity before and after the dying and the dyeing area are determined to evaluate the development quality of the cuticles of the apple fruits. The method can reduce artificial factor errors, is accurate and objective, and is a scientific means for evaluating the development situation of the cuticles of the mature apple fruits in production.

The invention provides a new method for screening and identifying genetic breeding new species of edible fungi, which solves the problems of obvious screening mark shortage, a waste of time and heavy work load during identifying whether the edible fungi or the new species contains manganese peroxidase or lignin peroxidase or not and slow qualitative identification and monitoring in the prior art. The new method comprises the following steps of: using aniline blue as one of materials for preparing a solid culture medium; inoculating a strain of the edible fungi to the solid culture medium; viewing the color change condition of the aniline blue solid culture medium after culturing; and identifying whether the edible fungi contains the manganese peroxidase or the lignin peroxidase or the manganese peroxidase and the lignin peroxidase or not. The new method can directly, conveniently and flexibly qualitatively judge whether the edible fungi contains the manganese peroxidase or the lignin peroxidase or not, is especially suitable for qualitatively detect two different types of wood-rotting fungi or new fused strains of other fungi and has low detection cost.

The invention discloses a method for staining and identifying arterial wall components. The method is characterized by comprising the following steps: sequentially performing heating, dewaxing, hydrating, hematoxylin iron staining, acid aniline blue staining and acid red staining on arterial sections of paraffin, dehydrating, clarifying and sealing, and enabling different components in the arterial walls to present different colors. According to the method disclosed by the invention, the cell nucleus is stained into dark blue or black to form dots or small strips; elastic fibers are stained into black and are rope-shaped; collagen fibers are stained into blue and are rope-shaped; endothelial cell cytoplasm is stained to be rosy to form strips; and smooth muscle cell cytoplasm is stained tobe rosy to form long shuttles. According to the stained colors and morphological features, the arterial wall components are identified. Different components of the arterial walls present different colors by virtue of staining, the arterial wall components are identified by distinguishing the colors and combining morphological changes, and arterial diagnosis and treatment can be assisted.

The invention relates to a method for separating Aspergillus tubingensis and application of Aspergillus tubingensis, straws are taken as a carrier to extract microorganisms capable of degrading straws, strains can be massively propagated through culture, red brown to brown black conidia are obtained through separation, the conidia are spherical or radial, most of molecular sporophores are generated from a matrix, and most of the molecular sporophores grow from the matrix. Most spore production structures are double layers, and bacterial colonies of top sacs are densely generated; the method comprises the following steps: selecting bacterial colonies, activating the selected bacterial colonies, culturing the bacterial colonies by using an ABTS chromogenic solid culture medium and an aniline blue chromogenic solid culture medium, seeing that decolorized rings are generated after the bacterial strains are dyed, proving that the bacterial strains can secrete laccase and lignin peroxidase, amplifying ITS sequences of the screened bacterial strains, and comparing the sequences in NCBI (National Center of Biotechnology Information) to obtain similar sequence information. A phylogenetic tree is established by utilizing MEGA-X, the strain is determined to be Aspergillus tubingensis, and the Aspergillus tubingensis can be applied to lignin degradation and dye decoloration.

The invention discloses a method for detecting generation of plant immunoreaction. The existing purification expression through prokaryotic expression products and the callose and reactive oxygen observation method are remained the same; the prokaryotic expression products of rice blast mycoprotein genes are adopted; 24 rice single-gene varieties containing different resistance genes, root organizations and sheaths of rice are processed according to the concentration of 5.0 mg/ml; the DAB dyeing and the aniline blue dyeing are implemented after 24 hours; the forming conditions of callose and reactive oxygen are directly observed through a fluorescence microscope; and the generation of the plant immunoreaction is judged through the forming conditions of the callose and the reactive oxygen. The method uses the prokaryotic expression products of purified effect protein genes to process the rice root organizations, and observes the generation of the callose and the reactive oxygen of the root organizations through the fluorescence microscope, so that the generation condition of the rice immunoreaction can be directly obtained, and the rice blast mycoprotein capable of leading a plurality of rice single-gene varieties containing different resistance genes and susceptible varieties to generate the immunoreaction is synchronously obtained.

The invention discloses acidic lipoxygenase and a preparation method and application thereof. A novel lipoxygenase gene MxLOX is obtained by cloning from an Myxococcus xanthus DK1622 strain genome, and the amino acid sequence of the novel lipoxygenase gene is shown as SEQ ID NO.2. The lipoxygenase has the highest activity under the conditions that the pH is 3.0 and the temperature is 30 DEG C, and has stable activity under the conditions that the pH is 2.5-5.0 and the temperature is 30 DEG C. Recombinant lipoxygenase rMxLOX can efficiently catalyze degradation of triphenylmethane dyes such as aniline blue and methyl blue, and has an potential application value in treatment of acidic industrial wastewater.

The invention belongs to the technical field of plant biology, and discloses an in-vitro identification method for gramineal self-incompatibility phenotype, which comprises the following steps: (1) inflorescence arrangement: the inflorescences of a paternal plant are pruned at the full-bloom stage, removing bloomed spikelets; (2) artificial flower forcing: the inflorescences are inserted into Hoagland nutrient solution, and flower forcing treatment is carried out under a completely dark condition for 8 to 12 hours; (3) in-vitro pistil culture: mature spikelets are chosen from the maternal inflorescences, and the pistils are intactly cut off under a stereoscopic microscope and inserted into 0.5 to 1 percent of agarose solid medium for in-vitro culture; (4) indoor pollination: in the morningof the next day, paternal pollen is used for pollinating the pistils; (5) pistil softening: the pollinated pistils are softened with 8 to 10mol/L of NaOH for 3 to 4 hours; (6) pistil dyeing: the softened pistils are dyed with 0.1 to 0.5 percent of aniline blue; (7) microscopic observation: the growth of the pollen tubes on the pistil stigmas are observed in an inverted fluorescence microscope. The method is simple and easy to master, the accuracy is high, and the method is suitable for being applied in the genetic analysis and crossbreeding research of miscanthus.