32 results about "Fluorescent stain method" patented technology

The invention relates to a fluorescent dyeing method for observing the microstructure of plants, which can effectively solve the problem of rapid film production and observation of the microstructure of plants. , the plant material was fixed in paraformaldehyde or methanol with a mass concentration of 4% configured by pH 8.0 PBS buffer solution for 20-40min, rinsed with pH 8.0 PBS buffer solution for 10-20min, and prepared by DMSO, Triton X-100 and fluorescent whitening agent 220 make staining solution, stain at room temperature for 10-60min, then rinse with PBS buffer for 10-15min, add DAPI solution dropwise for fluorescent staining of nucleic acid in cells, rinse with distilled water, and then add dropwise Glycerin, add a cover glass, observe and take pictures with a laser confocal microscope, the method of the present invention is simple, does not need a microtome, and the production time is short, and the three-dimensional structure of the plant can be observed, which is suitable for the observation of cells or tissues of various plants, clear , accurate, low cost and good effect.

The invention relates to a fluorescence dye for detecting fungus and dermatozoon. The technical problems that three dyeing liquid ingredients of a current fluorescence dye method cannot stably coexist for a long time, and need to be prepared and stored respectively, operating is tedious, the dyeing time is long, limitation exists in detected samples, some samples with the small impurities can only be dissolved, and the clinical application range is limited are solved. The fluorescence dye is prepared from water, a fluorescent whitening agent, alkali, a background redyeing agent, a chaotropic agent and a moisturizing agent. As the fluorescence dye is a single dosage form, the fluorescent whitening agent, strong alkali and the background redyeing agent can stably coexist; the stability of a product is good, dyeing can be completed only through one step, clinical using is more convenient and fast, original detection which can be completed in 5 minutes or above can be completed in tens of seconds through the fluorescence dye, and the detection efficiency of the fungus and the dermatozoon is greatly improved.

The invention discloses a fluorescent staining method for a cotton fiber. The method comprises the specific steps of pretreatment, modification, staining and fixation, wherein in the pretreatment step, a cloth sample is pretreated; in the modification step, water is added and then a modifier is added to the pretreated cloth sample to ensure that original negative charge on the surface of the cotton fiber is changed to positive charge, operation is conducted for 5min, temperature is raised to 70 DEG C at 1.5 DEG C per minute and kept for 30min, the cloth sample is taken out of a cylinder and washed with water once, and the modification is finished; in the staining step, water is added and then fluorescent paint for printing is added according to a color formula to the modified cloth sample, operation is conducted for 5min, temperature is raised to 70 DEG C at 1.5 DEG C per minute and kept for 30min, the cloth sample is taken out of the cylinder and washed with the water once, and the staining is finished; and in the fixation step, water is added and then an adhesive is added to the stained cloth sample, temperature rise and heat preservation are conducted, and the cloth sample is taken out of the cylinder, washed with the water cleanly, softened and dried. According to the fluorescent staining method for the cotton fiber, the fluorescent paint for the printing can be used for staining, and the hand feeling, fastness and cloth cover uniformity are improved.

The invention provides a fluorescent staining method of evaluating sperm motility of a tree shrew. The fluorescent staining method comprises the following steps: (1) preparing a TALP-HEPES sperm rinse; (2) collecting sperms; (3) washing the sperms; (4) staining; and (5) detecting by a microscope. According to the fluorescent staining method provided by the invention, by utilization of the characteristics that Hoeches 333422 and PI are different in reaction to dead and alive sperms and can be excited by a laser with a same wavelength, fluorescent staining is carried out on sperms of the tree shrew, and the sperm motility can be accurately detected without needing to switch a light source; in addition, as the two dyes respectively emit red and blue fluorescence after being excited by the laser, a dye which emits green fluorescence can also be introduced to detect the integrity of sperm acrosomes while the motility is detected, and the sperm motility and the integrity of the sperm acrosomes of the tree shrew are accurately detected by virtue of a fluorescence microscope or a flow cytometer.

The invention discloses a fluorescent staining method for evaluating sperm motility of domesticated mammals. The fluorescent staining method comprises the following step: according to the characteristic that a mammal live sperm probe Hoechst 333422 and a mammal dead sperm probe PI can be excited by the same ultraviolet light to emit fluorescent light, performing fluorescent staining on sperms of four important domesticated mammals which are pigs, cows, sheep and dogs to accurately perform sperm mobility evaluation. Compared with the conventional PI / SYBR-14 staining method, the PI / Hoechst 33342 staining method adopts Hoechst 33342 to substitute an expensive SYBR-14 probe, so that the detecting cost is greatly lowered. The fluorescent staining method can be used for successfully evaluating the sperm mobility of important domesticated mammals such as pigs, cows, sheep and dogs, and can be widely applied to evaluating the sperm motility of various domesticated mammals.

The invention relates to a acclimation direction adopted recycling small amount aborigine PHA high yield composition mushroom to do í«gage-restrictingí» acclimation primary activity sludge, force PHA composition mushroom to form dominance flora, and increase activity sludge composition PHA yield. Its purpose is to set aborigine PHA high yield composition recycling method, supply a simple activity sludge acclimation setting. Its operation steps are adopting Nile-Red fluorescent staining method to separate the PHA composition mushroom from the activity sludge; scalping PHA high yield mushroom as the recycling mushroom; its 0.1-0.25g are inoculate in reaction barrel to finish acclimation periods; discharging acclimation fluid; adding fermentation medium and microelement to do PHA ferment; collecting sludge deposition, drying, using chloroform to extract PHA; condensing extracted fluid in methanol to cement out PHA. This method can effectively increase activity sludge composition PHA yield from thirty percent to fifty percent.

The difficult problem of the contamination of cell cultured mycoplasma and the infection diagnosis of various mycoplasma urgently need a reliable and effective kit. Mycoplasma culture by a fluorescent staining method is reliable but takes time and trouble, and culture pollution is intensified. A direct PRC method for amplifying a mycoplasma gene, which is used for the routine detection of the mycoplasma is easy for cross contamination, and high false masculinity is only used as an auxiliary reference. The invention relates to a mycoplasma conservative gene which across links a plant arabidopsis LEY sequence into a reporting gene DNA by a hybridization probe and amplifies the indirect detection of a reporting gene. The mycoplasma conservative gene is characterized in that mycoplasma 16sRNA is hybridized with the head part and the tail part of the LEY reporting gene DNA whose head part and tail part are provided with mycoplasma conservative sequence probes; the reporting gene DNA is catalyzed into a ring by heat-proof ligase, and the annular reporting gene is reversely amplified by PCR so as to indirectly reflect the detection of the mycoplasma; by adjusting ligase reaction, the reporting gene is difficultly across linked by the cross contamination, thereby reducing the false masculinity; and if a system is contaminated, the reporting gene can be changed.

The invention discloses a fast fluorescent staining method of cruciferae crop pollen. The method comprises the following steps of S1, staining liquid preparation: firstly, dissolving Hoechst33258 fluorescent dye and cane sugar into a PBS (phosphate buffer solution); preparing a first solution with the Hoechst33258 concentration being 20 to 40mug / L and the cane sugar concentration being 8 to 10 weight percent; then, preparing a second solution containing 60 to 70 percent (v / v) of ethanol, 10 percent (v / v) of ethyl acetate and 30 to 20 percent (v / v) of glycerol; mixing and uniformly mixing the first solution and the second solution according to a volume ratio of 1:1 to obtain staining liquid; performing low-temperature light avoiding storage; S2, staining: taking staining liquid prepared inthe steps S1 on a glass slide; then, picking blossoming flowers or flower buds to be stained and observed from the inflorescence; picking the anther by tweezers to be directly put into staining liquidon the glass slide; slightly pinching the anther; spreading out pollen; taking out sundries; performing smearing so that the pollen and the staining liquid are uniformly mixed; then, cover glass is covered. The problems of complexity, time waste and labor waste of the existing pollen fluorescent staining method can be solved; meanwhile, the stability and the repeatability of the fluorescent staining method are improved.

The invention relates to a fluorescent staining method for separating dairy cow sperms, comprising the following steps of: centrifuging dairy cow semen before staining to remove part of seminal plasma, and mixing the semen after part of the seminal plasma is removed with fluorescent dye according to a certain proportion for staining. The novel fluorescent staining method has the advantages of good staining effect, great fluorescence intensity difference with X / Y sperms after staining, easy separation of sperms and the like, thereby not only improving the separating effect and efficiency of sperms, but also improving the use ratio of breeding oxen.

The invention discloses a fluorescent staining method of cuttlefish embryo development and nuclear behavior observation. The method comprises the following steps: (1) selecting a cuttlefish egg, peeling off outer egg membranes and reserving an innermost egg membrane; (2) submerging the cuttlefish egg treated in the step (1) in a fixing liquid for conservation at 4 DEG C, and changing the fixing liquid for twice to four times during the conservation period; (3) washing the cuttlefish egg fixed in the step (2) with buffer solution for twice or thrice, and staining with a fluorescent staining agent in dark environment; and (4) cutting off a capitulum at the animal pole of the cuttlefish egg treated in the step (3), laying on a slide glass, and imaging under a fluorescence microscope. The method has the advantages of not destroying substances and structures on the surface of fertilized eggs, is characterized by simple step and convenient operation, and can rapidly, clearly and directly observe embryonic development and nuclear behavior of the cuttlefish under the fluorescence microscope.

The invention belongs to the technical field of biotechnology and especially relates to a method for establishing a cell model for diabetes complicated with depression. The method comprises the following steps: adding glucose with a final concentration of 50 to 200 mmol / L and corticosterone with a concentration of 100 to 400 [mu]mol / L into a medium for culturing hippocampal neurons and carrying out intervention culture on the hippocampal neurons for 18 to 24 h so as to obtain the cell model. The cell model is evaluated through general morphological observation, HE dyeing, glucose consumption detection, intracellular neurotransmitter detection, MTT-process detection, flow cytometry, Hoechst fluorescent staining and TUNEL staining; and evaluation results show that the cell model has good effect and can be used for pathological and physiological research on diabetes complicated with depression, research on pathogenesis, in-vitro drug screening or drug evaluation.

The invention provides an immunofluorecence technique based method for evaluating activities of cryptosporidium parvum and giardia. The method has the following beneficial effects: the accurate, quick, simple, convenient and low-cost high throughput immunofluorecence assay technique is established and is used for evaluating the activities of cryptosporidium parvum and giardia in the ultraviolet (UV) disinfectant fluid; evaluation of the activities of cryptosporidium parvum and giardia in the UV irradiated water is realized by mainly carrying out competitive immunobinding on anti-pyrimidine-dimer (TDs) monoclonal antibodies and the TDs, taking fluorescein isothiocyanate as a marker and immunoglobulin (IgG) as a fluorescent substrate and binding 4',6-diamidino-2-phenylindole, dihydrochloride (DAPI); compared with the fluorescent staining method for evaluating the effect of UV on inactivating cryptosporidium parvum and giardia, the method provided by the invention has such excellent characteristics as strong specificity, high sensitivity and low professional operation requirements; and the cryptosporidium parvum and giardia activity evaluation method system is perfected by binding the method provided by the invention with the fluorescent staining method and other methods, and the method provided by the invention can be widely applied to the fields such as water quality monitoring, disease control and epidemic prevention and the like.

The invention discloses a single ultraviolet light excited fluorescent staining method used for evaluating sperm activity of Kunming wolf dogs. The method is realized by virtue of four steps of preparing a TCG sperm washing solution, washing sperms, staining and detecting with a microscope. According to the method disclosed by the invention, two types of fluorescent dyes which can be excited by ultraviolet light of a same wavelength are adopted to perform fluorescent staining on sperms of Kunming wolf dogs, and the sperm activity can be accurately identified without switching a light source; and moreover, one of the two types of fluorescent dyes emits red fluorescent light and the other one emits blue fluorescent light after laser excitation, so that the activity is detected, a dye emitting green fluorescent light can be introduced to detect acrosome integrality. The fluorescent staining method disclosed by the invention is simple to operate, accurate in result and high in reliability, and the sperm activity of the Kunming wolf dogs and the acrosome integrality can be accurately detected by using a fluorescence microscope or a flow cytometry.

The invention discloses a primer pair for identifying Scolopendra subspinipes multilans(L.)Koch and an application of the primer pair. The primer pair disclosed by the invention for identifying or assistant-identifying Scolopendra subspinipes multilans(L.)Koch is specifically the primer pair of two single-chain DNA molecules shown in sequences 1 and 2 in a sequence table. Experiments verify that by adopting the primers disclosed by the invention, Scolopendra subspinipes multilans(L.)Koch and Scolopendra multidens Newport are quickly and accurately identified by virtue of a quick PCR method and a fluorescent staining method and a fluorescent dye method is introduced to detect a true and false identification result, so that a technical support for field utilization on medicinal material molecular identification is realized.

The invention relates to a fluorescent staining method for separating dairy cow sperms, comprising the following steps of: centrifuging dairy cow semen before staining to remove part of seminal plasma, and mixing the semen after part of the seminal plasma is removed with fluorescent dye according to a certain proportion for staining. The novel fluorescent staining method has the advantages of good staining effect, great fluorescence intensity difference with X / Y sperms after staining, easy separation of sperms and the like, thereby not only improving the separating effect and efficiency of sperms, but also improving the use ratio of breeding oxen.

The invention discloses a pig oocyte membrane protein NHE1 fluorescent staining method, and relates to the technical field of cell biology. In order to more visually observe the period of cells and the co-localization condition of membrane protein NHE1 and microfilaments, the invention provides a porcine oocyte membrane protein NHE1 fluorescent staining method which comprises the following steps:extracting a cumulus oocyte complex from sexually mature porcine follicles, culturing, obtaining oocytes in a foaming period GV, a primary maturation and division early-stage GVBD, a primary maturation and division medium-stage MI and a secondary maturation and division medium-stage MII, sequentially removing cumulus cells through hyaluronidase and removing zona pellucida through streptavidin, performing membrane permeation and sealing on pig oocytes in each period, performing antibody incubation, and dyeing nuclei to prepare a sealing sheet, and observing under a multichannel confocal microscope. The invention provides a new technical support for researching the molecular mechanism of the porcine oocyte membrane protein NHE1 and other membrane proteins.

The invention discloses a fast fluorescent staining method of cruciferae crop pollen. The method comprises the following steps of S1, staining liquid preparation: firstly, dissolving Hoechst33258 fluorescent dye and cane sugar into a PBS (phosphate buffer solution); preparing a first solution with the Hoechst33258 concentration being 20 to 40mug / L and the cane sugar concentration being 8 to 10 weight percent; then, preparing a second solution containing 60 to 70 percent (v / v) of ethanol, 10 percent (v / v) of ethyl acetate and 30 to 20 percent (v / v) of glycerol; mixing and uniformly mixing the first solution and the second solution according to a volume ratio of 1:1 to obtain staining liquid; performing low-temperature light avoiding storage; S2, staining: taking staining liquid prepared inthe steps S1 on a glass slide; then, picking blossoming flowers or flower buds to be stained and observed from the inflorescence; picking the anther by tweezers to be directly put into staining liquidon the glass slide; slightly pinching the anther; spreading out pollen; taking out sundries; performing smearing so that the pollen and the staining liquid are uniformly mixed; then, cover glass is covered. The problems of complexity, time waste and labor waste of the existing pollen fluorescent staining method can be solved; meanwhile, the stability and the repeatability of the fluorescent staining method are improved.

The invention belongs to the technical field of plant virus detection, and specifically discloses a RT-LAMP kit and method for rapid detection of a sugarcane streak mosaic virus. The kit comprises a RT-LAMP primer group for detecting the sugarcane streak mosaic virus. The RT-LAMP primer group comprises a pair of outer primers (F3, B3) and a pair of inner primers (FIP, BIP). The nucleotide sequences of F3, B3, FIP, and BIP are represented by SEQ ID No.1-No.4. The detection method comprises following steps: extracting total nucleic acids of a sample to be detected, carrying out RT-LAMP reactionsby using the abovementioned kit, and determining whether the sample is infected by the sugarcane streak mosaic virus or not by a fluorescent staining method. The provided kit has the advantages of high specificity and high sensitivity and can accurately identify plants that are infected by the sugarcane streak mosaic virus; the operation of the detection method is simple and convenient; and the kit is suitable for rapid field detection of sugarcane streak mosaic virus.

The invention discloses a wheat root cell nucleus extraction method applicable to immunofluorescence analysis. The wheat root cell nucleus extraction method comprises the following steps: (1) obtaining wheat young roots; (2) extracting cell nucleuses; (3) detecting the cell nucleuses through a DAPI (Diamidino-phenyl-indole) fluorescent staining method; (4) carrying out immunofluorescence histochemical analysis; (5) carrying out statistic analysis. According to the cell nucleus extraction method provided by the invention, the time for extracting the cell nucleuses can be greatly reduced and theactivity of protein in the cell nucleuses is kept for a longer time; the accuracy and success rate of subsequent target protein detection are improved, the concentration of impurities in a cell nucleus suspension solution is reduced, influences, caused by the impurities, in an immunofluorescence assay are reduced, the quantity of the cell nucleuses in the cell nucleus suspension solution is increased and related statistical experiment and analysis are convenient to carry out subsequently. The extraction method provided by the invention is applicable to production of the cell nucleus suspension solution so that subsequent submicroscopic-level cytological related experiments are convenient to carry out.

The invention discloses a fluorescein carbon dot dyeing reagent for fungus detection. The fluorescein carbon dot dyeing reagent is prepared from the following components in percentage by mass: 0.01 to 0.1 percent of a fluorescein carbon dot material and 99.90 to 99.99 percent of water, wherein the fluorescein carbon dot material is prepared from the following raw materials in percentage by mass: 1%-2% of a fluorescent whitening agent, 10%-15% of citric acid, 8%-12% of ethylenediamine and 73%-78% of water. The reagent contains the fluorescein carbon dot material, and the fluorescein carbon dot material is stable in fluorescence, high in fluorescence intensity and good in fungus imaging effect. The invention further discloses a dyeing method adopting the fluorescein carbon dot dyeing reagent. The method is simple, easy to implement and short in consumed time. The invention also discloses application of the fluorescein carbon dot staining reagent in fungus detection by adopting a fluorescent staining method.

The invention discloses a fluorescent staining method for evaluating sperm motility of domesticated mammals. The fluorescent staining method comprises the following step: according to the characteristic that a mammal live sperm probe Hoechst 333422 and a mammal dead sperm probe PI can be excited by the same ultraviolet light to emit fluorescent light, performing fluorescent staining on sperms of four important domesticated mammals which are pigs, cows, sheep and dogs to accurately perform sperm mobility evaluation. Compared with the conventional PI / SYBR-14 staining method, the PI / Hoechst 33342 staining method adopts Hoechst 33342 to substitute an expensive SYBR-14 probe, so that the detecting cost is greatly lowered. The fluorescent staining method can be used for successfully evaluating the sperm mobility of important domesticated mammals such as pigs, cows, sheep and dogs, and can be widely applied to evaluating the sperm motility of various domesticated mammals.

The invention discloses a primer pair for identifying radish seeds and an application of the primer pair. The primer pair for identifying or helping to identify the radish seeds is specifically composed of two single-chain DNA molecules as shown in a sequence 1 and a sequence 2 in the sequence table. Experiments prove that the primers are adopted to realize quick and accurate identification of radish seeds, Chinese dodder seeds, perilla fruits, white mustard seeds, feather cockscomb seeds, flastem milkvetch seeds, seeds of garden balsam, chingma abutilon seeds, rapeseeds and the like are identified by use of a fast PCR method and a fluorescent staining method; and a fluorescent dye method is introduced to detect the true and false identification results, and technical support is provided for the field application of molecular identification of medicinal materials.

The invention provides a fluorescent staining method for living cell imaging, cells are subjected to double staining through the first fluorescent biomarker and the second fluorescent biomarker, clear fluorescent cell images are detected and displayed through a fluorescence microscope, and according to the obtained images, the cell morphology can be observed while the cell nucleus morphology can be observed.

The invention discloses a spatial transcriptome data processing method and device and a computer readable storage medium. The method comprises the following steps: acquiring a Hamp of a target sample slice through a space transcriptome technology; e, dyeing an image and a space transcriptome sequencing analysis result; performing fluorescence-labeled antibody staining of the target protein on the target sample section through a fluorescence staining method to obtain a fluorescence staining image of the target protein; the method comprises the following steps: performing Hamp; the E dyeing image and the fluorescent dyeing image are superposed to obtain a superposed image; extracting site information corresponding to a fluorescence positive signal in the superposed image; and analyzing and processing the transcriptome data corresponding to the site information according to the spatial transcriptome sequencing analysis result to obtain a spatial transcriptome data analysis result of the target sample slice at the site information. According to the method and the device, the technical problem that the reliability is relatively low as only a space transcriptome technology combined with single morphological information is adopted to analyze the sample in the related technology is solved.

The present invention relates to fluorescent premix particles, a fluorescent stain containing the same, and a fluorescent staining method in which these are used, and the fluorescent premix particles are particles including: phosphor integrated particles that are modified with a first reactive substance; and at least one target protein-binding substance that is modified with a second reactive substance and is selected from the group consisting of an antibody and an aptamer, wherein the phosphor integrated particles and the at least one target protein-binding substance are linked by interaction between the first reactive substance and the second reactive substance.

The invention discloses application of pterostilbene in inhibiting vascular endothelial apoptosis. The effective concentration of the pterostilbene in application of inhibiting vascular endothelial apoptosis is between 0.5 and 5 mu M. The function of the pterostilbene in inhibiting vascular endothelial apoptosis is sufficiently proved in a cell morphological alteration experiment observed by an inverted phase contrast microscope, a nucleus condensation and karyorrhexis situation experiment observed by a laser scanning confocal microscope, a nucleus heterochromatin agglutination situation experiment observed by a Tunel fluorescence microscope, a dye marked apoptotic cell experiment, an activity experiment of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay detectedcell succinodehydrogenase, an affecting experiment of the pterostilbene to apoE- / - mice aortic vascular endothelial apoptosis detected by a frozen section immunofluorescence staining method, a JC-1 dyeing detected mitochondrial membrane potential change experiment and a Westernblot detected apoptosis related protein expression experiment. The pterostilbene has remarkable effect in inhibiting vascular endothelial apoptosis, so that an attractive prospect is provided to the development of preparing a medicament for inhibiting vascular endothelial apoptosis and a novel cardiovascular medicament.

The invention relates to a fluorescent staining method for plasmodium detection and a reagent thereof. The method is characterized by comprising the following steps: obtaining a small molecule compound capable of being specifically combined with a plasmodium nuclear structure through a molecular fishing experiment, and modifying and marking the small molecule compound to form a specific fluorescein marked small molecule; by using the reagent provided by the invention, fluorescence staining detection on plasmodium in blood is realized; and after the reagent is mixed with a sample, the specific fluorescein labeled small molecules are combined with the plasmodium, so that the red blood cells containing the plasmodium are in bright color or specific color under a fluorescence microscope, and other blood cells are in black or background color.