40 results about "Nuclear staining" patented technology

The invention relates to yellow fluorescence carbon dots with high quantum yield. A preparation method of the yellow fluorescence carbon dots is characterized by comprising the following steps: dissolving organic acid and organic amine in water; performing microwave radiation after fully mixing the organic acid and the organic amine, thus obtaining a brown solid crude product; performing reverse-phase silica gel column chromatography purification by taking a mixed solution of the water and methyl alcohol as an eluting agent, thus obtaining the yellow fluorescence carbon dots. The yellow fluorescence carbon dots are characterized in that the average grain diameter is 3.0 to 4.0 nm; bright yellow fluorescent light can be emitted under the irradiation of an ultraviolet lamp, the excitation wavelength is 400 to 420 nm, the emission wavelength is 510 to 550 nm, the luminescent property of solids is shown, and the quantum yield can reach 44 percent. The yellow fluorescence carbon dots prepared by the invention are good in water solubility and stable in fluorescence signals, are suitable for cell imaging under physiological conditions, have a function of nuclear staining and can be widely applied to the fields of analysis and detection, biochemical sensing, biological imaging, biological marking and the like.

The invention provides a single cell separation method based on droplet micro-fluidic chips. The method concretely comprises the following steps: A, a single cell suspension flows into a dispersed phase inlet channel from a dispersed phase inlet; B, an oil phase liquid flows into a continuous phase inlet channel from a continuous phase inlet; C, the above two phases merge to form droplets enclosing single cells, the droplets flow over a droplet capturing unit with the generation of a large number of droplets in a liquid storage pool, and stand for 2-5 min, and the superfluous droplets are sucked out when the droplets slowly settle into the droplet capture unit; and D, the droplet chips which capture the single cells are cultured in a 37 DEG C incubator, then DAPI is added to carry out nuclear staining, and the single cell capture rate is detected. The method has the advantages of simplicity and rapidity in operation, small use amounts of the cells and reagents, low experiment cost, high integration and wide application range.

The present invention relates to an immunofluorescence kit for detecting PD-L1 and CD8 antigens and a detection method using the kit. The kit comprises the following reagents: buffer solutions CYP1, CYP2 and CYPP, a blocking solution, a specific antibody for detecting PD-L1, a corresponding fluorescent second antibody, a fluorescently labeled CD45 antibody, a fluorescently labeled CD8 antibody, anantibody dilution solution and a nuclear staining solution. The invention also provides a method for carrying out antigen detection by using the kit. A detection result of the kit can guide actual drug use, and not only can reflect the whole tumor comprising carcinoma in situ and possibly existent invisible micro-metastasis load, but also can reflect the expression condition of the PD-L1 in CTC;a reference can be provided for precise selection of therapeutic drugs according to the proportion of PD-L1 and CD8 positive cells in PBMC; and the kit is more convenient than a detection technology based on tumor tissues in material acquisition.

The invention is a method for analyzing immature reticulocytes for the presence of micronuclei. The method includes reticulocyte enrichment, fluorescent labeling, micronuclei staining, and analysis using single-laser flow cytometry. The invention also includes kits containing reagents to use in the method.

The invention discloses a breast cancer Ki67 / ER / PR nuclear staining cell counting method based on staining separation. The method comprises the steps that firstly, a breast cancer pathological sectionimage is dyed and separated, an image IH based on an H coloring agent and an image IDAB based on a DAB color developing agent are obtained, then the influence of intercellular cytoplasm is eliminatedthrough filtering, and an image IH, filterd, IDAB and filterd are obtained; secondly, counting the negative cells according to IH and filterd, counting strong positive, middle positive and weak positive cells according to IDAB, filterd and the brightness map VDAB of the IH and filterd, and finally obtaining the number of the negative cells, the number of the positive cells and the number of the positive cells (divided into strong positive, middle positive and weak positive) and the probability of occupying the total number of the cells. According to the breast cancer nuclear staining cell counting method based on staining separation, cell counting can be rapidly and effectively achieved, doctors are assisted in work, the workload of the doctors is reduced, and meanwhile the calculation accuracy is guaranteed.

The invention discloses a floating sphere immunofluorescent staining method and a staining device, belonging to methods for coloring a sample for test. The staining method comprises the following steps: separation of spheres from a culture medium, fixing by stationary liquid, adding of Triton X-100 transparent cells, closing by confining liquid, adding of an interest protein-resisting primary antibody working solution, adding of a primary antibody-resisting fluorescent secondary antibody working solution, and cell nucleus staining by DAPI dye liquor. The staining device comprises a cell chamber and a staining sleeve part arranged at the outer part of the lower end of the cell chamber in a sleeving way. The floating sphere immunofluorescent staining method has the advantages of high efficiency, simpleness, convenience and the like, an antibody usage amount is reduced, experiment cost is reduced, manpower and time are saved, an accurate and reliable experiment result is obtained on the basis that a fundamental principle that immunofluorescent staining is not changed and specially training experimenters is not needed, sphere loss in an experiment operation process is avoided, an experiment success rate is increased, and the efficiency is improved.

The present invention relates to compositions, methods, and kits suitable for detecting nucleic acids in a biological sample. The nuclear staining composition of the present invention contains a pH buffering reagent, a solubilizing reagent, a basic dye, and an aqueous medium. The composition can be used alone to detect nucleic acids in a biological sample or in combination with other histological dyes for nuclear counterstaining.

The invention relates to a full-automatic liquid-based cell tableting and staining integrated machine, comprising: a mounting frame, a circulating sample injection mechanism, a specimen liquid puncture-adding mechanism, a slide loading mechanism, a code printing mechanism, an alcohol fixing and nuclear staining mechanism, a cytoplasm staining mechanism, a rinsing mechanism, a slide collecting mechanism and a slide clamp circulating mechanism, wherein the mounting frame is the skeleton of the whole machine; the slide clamp circulating mechanism is used for circulating a slide clamp in the machine; the circulating sample injection mechanism is used for transmitting a sample to the specimen liquid puncture-adding mechanism; the specimen liquid puncture-adding mechanism is used for adding thesample liquid to the slide clamp; the slide loading mechanism is used for loading a slide into the slide clamp; the code printing mechanism is used for printing a code on the slide in the slide clamp;the alcohol fixing and nuclear staining mechanism is used for carrying out fixing and nuclear staining on the sample; the cytoplasm staining mechanism is used for carrying out cytoplasm staining on the sample; the rinsing mechanism is used for washing floating color of the sample; and the slide collecting mechanism is used for taking out the slide from the slide clamp and putting into a collection box. The machine is easy to operate, high in automation degree and high in efficiency.

A method is provided for improving the shelf-life of a nuclear staining solution. In particular, the present invention provides a method of adding an antioxidant to a hematoxylin staining solution which maintains the performance of the stain over its shelf-life.

The invention provides a fluorescent carbon dot for nuclear staining. The fluorescent carbon dot is prepared from the following steps: 1) dissolving folic acid and m-phenylenediamine in water, and carrying out heating for a reaction; and 2) after the reaction, performing centrifuging and silica gel column chromatography, collecting a yellow-green fluorescent part, transferring the yellow-green fluorescent part into water after rotary evaporation and then carrying out lyophilization. The carbon dot prepared in the invention emits green light under ultraviolet excitation, has large Stokes shift,is consistent with the existing commercial 405 laser at the optimal excitation position, and can be well applied to a laser confocal microscope.

The invention provides a method for diagnosing a melanocytic proliferation in a subject comprising staining a sample of lesional melanocytes with an antibody against soluble adenylyl cyclase (sAC) and interpreting the sAC staining pattern, which is associated with a diagnosis of a melanocytic proliferation. The sAC staining pattern, which is complex, is discriminatory and distinctive according to the nature of the melanocytic proliferation. The sAC staining pattern comprises one or more of dot-like Golgi staining, broad granular Golgi staining, diffuse cytoplasmic staining, nucleolar staining, incomplete granular nuclear staining, and pan-nuclear staining. The method of the invention is particularly useful in confirming or disaffirming a diagnosis reached through conventional histologic examination of a sample. Additionally, the invention provides a kit for use in interpreting melanocytic proliferations.

The invention discloses a cancer cell recognition and diagnosis system, which comprises a cell image preprocessing module for selecting proper structural elements by adopting an improved mathematicalmorphology opening and closing filtering algorithm to remove background impurities and small normal discrete cells of a cell image; a cell image segmentation module used for extracting a cell nucleusregion by adopting an improved region growth algorithm, extracting a non-laminated cell body contour by adopting a watershed algorithm based on price tag control, and extracting a laminated cell contour by adopting a segmentation algorithm based on a snappe model; a cancer cell image feature extraction module used for calculating the nuclear-to-mass ratio of each region after segmentation, whetherthe cell nucleus size is uniform, whether nuclear staining is abnormal and whether the nucleus spacing is uniform. and a cell image classification and recognition module which recognizes cancer cellsthrough a neural network recognition technology. The system can be used for efficiently and accurately carrying out quantitative analysis and detection identification on cell images.

The invention discloses a method for marking germ cells and Sertoli cells of spermatogenic epithelium of turbot in different developmental stages and an application. Fragment Amh / Sox9 / Gsdf is amplified from cDNA of turbot spermary, cloning, preferring and plasmid extraction are performed sequentially, plasmids are linearized by restriction enzymes, probes are prepared after plasmid recovery, fixation, dehydration and entrapment are performed after sample processing, fluorescence in situ hybridization is performed after slicing, finally, nuclear staining is performed, and slice sealing, observation and photographing are performed. The method has the beneficial effects as follows: spermatids and the Sertoli cells of the spermatogenic epithelium of the turbot in different developmental stagesare separated out skillfully, and the marking method is simple and feasible and higher in precision; the method for marking the germ cells and the Sertoli cells of the spermatogenic epithelium of theturbot in different developmental stages can be applied to distinguishing and marking of cells of the spermatogenic epithelium of other marine organisms, and an effective method and a scientific basis are provided for studying spermatogenesis of fish and environment in the spermatogenic epithelium in future.

A method is provided for improving the shelf-life of a nuclear staining solution. In particular, the present invention provides a method of adding an antioxidant to a hematoxylin staining solution which maintains the performance of the stain over its shelf-life.

This invention provides isolated nucleic acid and amino acid sequences encoding VG5Q, a novel angiogenic growth factor protein with pro-angiogenic activity, a forkhead-associated domain, a G-patch domain; characteristic subcellular localization in an in vitro Matrigel model of angiogenesis: towards the cell periphery in early stages of tubulogenesis, between cells in newly formed endothelial tubes, and no nuclear staining after 24 hours; is expressed in endothelial cells; is secreted during angiogenesis; and interacts with TWEAK. The invention also provides for expression vectors containing nucleic acid sequences encoding VG5Q protein, and host cells containing one or more expression vectors for the recombinant expression of VG5Q. The invention also provides for methods of using VG5Q for the diagnosis and treatment of angiogenesis-mediated diseases or disorders.

The invention relates to a nuclear staining cell counting method based on deep learning of an incomplete label, computer equipment and a storage medium, and the method comprises the following steps: (1) making label data: loading a pathological image to label software, and obtaining sub-mask image data pairs of all sub-images and positive cells; (2) model training: training a convolutional neural network model to respectively obtain a trained positive cell convolutional neural network model and a trained negative cell convolutional neural network model; (3) reasoning stage: inputting pathological images to be detected into the trained convolutional neural network model to obtain real mask images; and (4) post-treatment stage: calculating the number of positive cells and negative cells, and calculating to obtain the proportion p of the positive cells in all the cells. The method does not need extra parameters, is high in universality, greatly reduces manual adjustment, and effectively improves the recognition accuracy and robustness. The method is faster, more accurate and more effective in data marking.

A kit is disclosed for preparing a measurement sample for detecting live cells, injured cells, VNC cells and dead microorganism cells in a test sample by the following steps:a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample,b) the step of treating the test sample with a topoisomerase poison and / or a DNA gyrase poison.c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, andd) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

The invention discloses a Pap staining solution and application thereof, and belongs to the field of biotechnology. Specifically, the Pap staining solution contains a cell nuclear staining solution and a cytosolic staining solution. The Pap staining solution provided by the invention is simple to operate, and ensures uniform staining, clear cell structure after staining and bright cell colors, soas to facilitate diagnostic observation and judgment of clinical pathology.

The invention belongs to the technical field of image analysis, and discloses a DNA fluorescence in situ hybridization BCR / ABL fusion state detection method and a detection system, which are characterized in that a CNN identification model is utilized to generate pseudonucleus staining of cells from a phase difference image, and FISH cell nucleuses are positioned and classified; and positioning and classifying each independent fluorescence signal, and dividing the number of color development points of the same classification in each photo by the total number of display points in the photo to obtain a BCR / ABL fusion ratio. The invention provides a system for detecting the fusion level of cell nucleuses and BCR / ABL genes by analyzing fluorescence in situ hybridization (FISH) images and calculating the image ratio of the number of abnormal cell nucleuses related to all classified cell nucleuses as an index for classifying the BCR / ABL gene fusion states of corresponding tumor samples. The detection method is high in detection efficiency, accurate in detection result and capable of achieving automatic detection.

The invention relates to an immunohistochemical nuclear staining section cell positioning multi-domain co-adaptation training method which is used for fully training a cell key point detection model under an only single-domain labeling data set. The method includes training the cell positioning model by adopting the source domain image and the target domain image, alternately inputting the sourcedomain image and the target domain image into an encoder to perform feature extraction, performing feature extraction on the source domain image to obtain a first feature, and performing feature extraction on the target domain image to obtain a second feature; inputting the first feature and the second feature into a discriminator for feature discrimination; when the loss function of the discriminator reaches a set condition, taking the extracted first feature and second feature as domain invariant features; alternately inputting the first feature and the second feature into a decoder for decoding, and performing activation operation to obtain a corresponding confidence map; in the training process, enabling the encoder and the decoder to perform parameter updating through continuous iteration; and when the number of training iterations reaches a specified number, ending the training.

The invention discloses a hybridization probe for detecting von hippel-lindau (VHL) gene large deletion and a detection method and a kit, which relate to the field of gene detection, wherein the hybridization probe is marked with fluorescence signals, and is prepared from probe primers with nucleotide sequences which is showed in sequence identifier number 1-6 (SEQ ID NO.1-6). The detection method comprises steps: cell smears are prepared from collected samples to be detected, hybridization probe is arranged in a hybridization solution to have a hybridization reaction with cell and obtain hybridization products, the hybridization products are washed, dyeing of cell nucleus are processed to obtain cell smears after nuclear staining, the cell smears after nuclear staining are observed through a fluorescence microscope, thereby judging whether deoxyribose nucleic acids (DNA) of the samples to be detected happen VHL gene large deletion. The kit comprises the hybridization probe. The hybridization probe is strong in specificity, high in sensitivity, simple in detection method and low in cost of the kit, and can rapidly and accurately detect VHL gene large deletion.

According to the following steps, live cells, injured cells, VNC cells and dead cells of a microorganism in a test sample are detected by flow cytometry:a) the step of treating the test sample with an enzyme having an activity of decomposing cells other than those of the microorganism, colloidal particles of proteins or lipids existing in the test sample,b) the step of treating the test sample with a topoisomerase poison and / or a DNA gyrase poison.c) the step of treating the test sample treated in the steps a) and b) with a nuclear stain agent, andd) the step of detecting the microorganism in the test sample treated with the nuclear stain agent by flow cytometry.

The invention discloses a cell nucleus DNA staining method. The method comprises the following steps: preparing an AF stationary liquid, a hydrolysate, a rinsing liquid and an eosin staining liquid; preparing a cell DNA staining solution which is composed of a cell DNA staining solution A and a cell DNA staining solution B in a volume ratio of 1: 1; placing a pathological sample into the AF stationary liquid to be fixed; washing the sample with running water, and then placing the sample into a hydrolysate to hydrolyze; washing the sample with running water, and then placing the sample in the cell DNA staining solution for staining; washing the sample with running water, and then placing the sample in a rinsing liquid for rinsing; and carrying out dehydrating with ethanol step by step, placing the sample into an eosin staining solution, and carrying out decolorizing with absolute ethyl alcohol, airing, and sealing. According to the method, dyeing time is greatly shortened, the hydrolysate is composed of non-precursor chemicals and belongs to an environment-friendly reagent, compared with a traditional stationary liquid, the AF stationary liquid does not include acetic acid and lesssimulates the human body, and the cell DNA staining solution is simple to prepare, does not need to be heated and boiled, has the validity period longer than 18 months, is ready-to-use and is suitablefor commercial popularization.

The invention discloses a method for nanotube-height-aided control of cytoskeleton change. The method comprises steps of performing anodization of pure titanium or low-modulus biphase titanium using an organic solvent; representing the microscopic structure of an anodization product using a microscopic means; preparing a titanium metal piece of nanopore and naotube microstructures; separating bone narrow mesenchymal stem cells, adhering three-generation cells on a surface-modified metal piece for incubation; observing cell adhesion effects by MTT, DYPI nuclear staining and SEM; placing the metal testing piece for cell incubation in culture holes of an aseptic elastic plate, and placing the metal testing piece in a universal testing system in a constant temperature state and providing the metal testing piece with different periods, frequencies and load compressive stress; and observing and testing cell deformation with a SEM electron microscope or optical microscope, and detecting representation of cell proliferation and differentiation. The method enables the elastic deformation signal of a modified titanium substrate surface to be amplified, thereby influencing osteoblast adhered on the modified titanium substrate surface to receive greater simulation, and leading to larger cytoskeleton change.

The invention discloses a method for preparing frozen sections of horse, donkey and mule testicles. The method comprises the following steps that after blocky testicle tissues are placed in a Bouin fixing solution to be fixed, the fixed testicle tissues are dehydrated in a sucrose solution, and the dehydrated testicle tissues are embedded, and frozen and sliced to obtain the testicle frozen sections. The method is used for preparing the frozen sections of the horse, donkey and mule testicles, and can denature, solidify and precipitate protein, and keep original morphological structures of cells and tissues to prevent autolysis and decay of the tissues, enzymes in the cells are converted into insoluble substances so as to keep the original structures, the tissues are in a certain hardened state after being fixed, the toughness of the tissues is improved, deformation is not easy, the cell integrity is kept, and the good cell tissue morphological structures can be shown after HE stainingand DAPI nuclear staining.

The invention discloses a spinal cord needle biopsy tissue fixation decalcification solution, a preparation method and a decalcification method. The spinal cord needle biopsy tissue fixation decalcification solution is prepared from the following components in parts by weight: 30-35 parts of formaldehyde, 300-400 parts of methanol, 400-500 parts of an ethylenediamine tetraacetic acid saturated solution, 7-9 parts of a PBS buffer solution, 25-35 parts of an accelerant, 1-3 parts of sodium chloride and 100-150 parts of water. The decalcification solution is small in tissue damage and better in tissue integrity, meanwhile, the decalcification time is greatly shortened, the decalcification time can be shortened from traditional 72 h to 24 h, the requirement for rapidly obtaining a pathological diagnosis report is greatly met, meanwhile, excimer detection can be conducted, accurate medication can be conducted on later-stage targeted therapy, the diagnosis efficiency is improved, and after decalcification treatment, the staining effect of a tissue specimen is better, the structure is clearer, the structures of soft tissues and bone groups in the section are completely displayed, the cell nucleus staining is clear, the nucleoplasm contrast is distinct, and the method has higher application value in the spinal cord needle biopsy tissue sheet preparation.

This invention provides isolated nucleic acid and amino acid sequences encoding VG5Q, a novel angiogenic growth factor protein with pro-angiogenic activity, a forkhead-associated domain, a G-patch domain; characteristic subcellular localization in an in vitro Matrigel model of angiogenesis: towards the cell periphery in early stages of tubulogenesis, between cells in newly formed endothelial tubes, and no nuclear staining after 24 hours; is expressed in endothelial cells; is secreted during angiogenesis; and interacts with TWEAK. The invention also provides for expression vectors containing nucleic acid sequences encoding VG5Q protein, and host cells containing one or more expression vectors for the recombinant expression of VG5Q. The invention also provides for methods of using VG5Q for the diagnosis and treatment of angiogenesis-mediated diseases or disorders.

The invention provides a subcellular localization kit for infecting tobacco and application of the subcellular localization kit. The subcellular localization kit comprises a buffer reagent A1, a reaction bacterial liquid A2 for reducing plant immunity, nuclear localization agrobacterium liquid A3 and agrobacterium competence A4. According to the kit, autoimmune response of plants is suppressed byusing plant resistance response profilin, the infection efficiency of agrobacterium is improved, and the expression of an exogenous gene is enhanced, therefore, a fluorescence signal is relatively strong and is observed more easily. Moreover, the kit contains nuclear localization control components, so that contact with toxic and cancerogenic substances in the process of using DAPI (Diamidino-Phenyl-Indole) nuclear staining is avoided; meanwhile, the test cost is reduced, and environmental pollution is reduced.

The invention discloses a preparation method of a nucleic acid fluorescence probe compound AzosD for nuclear staining. The preparation method is characterized by comprising steps as follows: 6-amino-2-methylquinoline is dissolved in acetonitrile, N-bromosuccinimide is added, a reaction is performed, and an intermediate 1 is obtained; the intermediate 1 is dissolved in a water, ethanol and toluenemixed solution, 3-(N,N-dimethylamino)phenylboronic acid, tetrakis(triphenylphosphine)palladium and sodium carbonate are added, refluxing reaction is performed under nitrogen protection, and an intermediate 2 is obtained; the intermediate 2 is dissolved by hydrochloric acid, sodium nitrite is added to the solution under the condition of ice-water bath, pH of the solution is regulated to be neutralby a sodium hydroxide aqueous solution, and 3-methyl-11-(N,N-dimethylamino) cinnoline[3,4-e]quinoline, namely AzosD, is obtained by separation and purification. The compound is good in photobleachingresistant effect, high in biocompatibility, high in enzyme-resistant capacity and high in nuclear targeting, can be applied to specific staining of nucleuses of living cells, and has broad applicationprospect in the fields of long-term monitoring of nuclear morphometry and observation for nuclear morphology change in the cellular physiological process.

The invention discloses a preparation method of a nucleic acid fluorescence probe compound AzosD for nuclear staining. The preparation method is characterized by comprising steps as follows: 6-amino-2-methylquinoline is dissolved in acetonitrile, N-bromosuccinimide is added, a reaction is performed, and an intermediate 1 is obtained; the intermediate 1 is dissolved in a water, ethanol and toluenemixed solution, 3-(N,N-dimethylamino)phenylboronic acid, tetrakis(triphenylphosphine)palladium and sodium carbonate are added, refluxing reaction is performed under nitrogen protection, and an intermediate 2 is obtained; the intermediate 2 is dissolved by hydrochloric acid, sodium nitrite is added to the solution under the condition of ice-water bath, pH of the solution is regulated to be neutralby a sodium hydroxide aqueous solution, and 3-methyl-11-(N,N-dimethylamino) cinnoline[3,4-e]quinoline, namely AzosD, is obtained by separation and purification. The compound is good in photobleachingresistant effect, high in biocompatibility, high in enzyme-resistant capacity and high in nuclear targeting, can be applied to specific staining of nucleuses of living cells, and has broad applicationprospect in the fields of long-term monitoring of nuclear morphometry and observation for nuclear morphology change in the cellular physiological process.