34 results about "Stain method" patented technology

This invention provides a PAM staining method that enables staining suitable for magnified observation of various samples using a range of microscopes, from optical microscopes to electron microscopes. [Solution] A PAM staining method comprising the steps of: silvering a biological sample; coloring the biological sample; and fixing the color of the biological sample, wherein the biological sample is a biologically derived sample that has been oxidized; the silvering step involves immersing the biological sample in a methenamine silver aqueous solution at room temperature, raising the methenamine silver aqueous solution from room temperature to 62°C or higher and 68°C or lower, and carrying out the silvering reaction for 30 to 40 minutes from the start of the reaction at room temperature; the coloring step involves immersing the silvered biological sample in a 0.2% gold chloride aqueous solution at room temperature for 8 to 12 hours; and the fixing step involves immersing the sample in a hard film fixing solution containing 11% to 13% sodium thiosulfate and aluminum potassium sulfate dodecahydrate for 3 minutes or more at room temperature.

The application discloses a kind of for carcinosarcoma evaluation section and its preparation method and application, belong to pathological technical field.The preparation method includes the following steps: reticular fiber staining: silver staining method is carried out to the reticular fiber in section tissue and is dyed;Immunohistochemical staining: AE1 / AE3 protein in section tissue is marked and dyed by immunohistochemical staining method.The section obtained by the preparation method can present two kinds of staining on a tissue section, improve the expression effect of cancer, sarcoma, carcinosarcoma, reduce the identification difficulty, and be beneficial to tumor accurate typing.

The invention discloses a method for predicting the curative effect of triple negative breast cancer immunotherapy by spatially quantizing CD39 + CD103 + CD8 + T cells, and relates to the technical field of biology.The method comprises the following specific steps that S1, an FFPE tumor tissue sample of a TNBC subject before immunotherapy is obtained, and a continuous section with the thickness of 3-5 microns is prepared and attached to a positive charge glass slide; s2, staining the section by using a combination containing CD8 / CD103 / CD39 / Pan-CK antibodies through a TSA sequential staining method, and then re-staining cell nucleuses by using DAPI; according to the method, spatial quantitative analysis is performed on a specific CD39 + CD103 + CD8 + T cell subset, a percentage index of target cells in CD8 + T cells and an average distance index of the target cells and nearest Pan-CK positive cells in a cancer nest region are set, and the target cells are determined according to comparison between the two indexes and a preset threshold value. According to the method, whether a subject is a potential benefit crowd treated by an anti-PD-1 / PD-L1 immune checkpoint inhibitor or not is accurately judged, a standardized technical means is provided for clinically screening the potential benefit crowd for immunotherapy, the accuracy and effectiveness of triple-negative breast cancer immunotherapy are effectively improved, and invalid treatment is reduced.

This invention discloses a high-throughput automated immunofluorescence staining device and its staining method. The device includes a liquid injection motion module, a reaction chamber module, a supply and waste collection module, and a control system. The liquid injection motion module includes an X-axis beam and a pipetting and auxiliary material robotic arm, which are driven in parallel to achieve sequential operation. The reaction chamber module includes a temperature-controlled station, a low-temperature reagent storage unit, and an auxiliary material supply unit. The supply and waste collection module includes a waste removal and cleaning unit and an auxiliary material waste collection unit. This invention achieves a 96-station array through a temperature-controlled reaction station, combined with the dual-drive parallel operation of the pipetting and auxiliary material robotic arms, eliminating action waiting time and significantly improving throughput. The use of a closed semiconductor cooling component linked with an automatic flip-top mechanism ensures uniform reagent activity over a long period. A plate-mounted small vacuum pump drives a rigid pipeline with deep-hole waste suction, combined with high-flow-rate wide-area spraying, to thoroughly remove waste liquid and reduce cross-contamination. This achieves full-process automation, significantly improving staining quality and efficiency.

Staining devices and methods for automated systems are provided herein. A dyeing method includes disposing a substrate proximate a dyed surface to establish an overlap between the dyed surface and the substrate. The staining method further includes maintaining a gap between the stained surface and the substrate to define a chromosome product according to the overlap and the gap, where the chromosome product has one or more meniscus edges. The staining method further includes disposing a portion of the staining fluid over the staining surface and distributing the staining fluid over the substrate.

The invention relates to a wine stain treatment system based on stain characteristics and dual-wavelength parameter matching, and the system comprises a data collection module which is used for collecting a wine stain region parameter, a patient skin parameter and a historical treatment parameter of a patient; presetting a dual-wavelength wine stain treatment method library; the color spot mapping module is used for extracting color spot characteristic parameters from the wine color spot area parameters and determining adaptive matching values of the color spot characteristic parameters and the dual-wavelength treatment methods so as to construct a characteristic mapping matrix; the matrix optimization and adjustment module is used for extracting a core influence factor and a weighting coefficient from the skin parameters of the patient and carrying out weighting operation and adjustment on the core influence factor and the weighting coefficient and the feature mapping matrix; the treatment method matching module is used for matching with a method library according to the feature mapping matrix; and the color spot treatment module is used for setting a dual-wavelength light source to perform color spot treatment according to the acquired dual-wavelength treatment method. Compared with the prior art, the method has the advantage that the safety and the personalized adaptability of a spot treatment scheme are improved.

A method for staining reticulocytes includes staining reagent A and stabilizing reagent B; stabilizing reagent B is used M seconds after staining reagent A is mixed with the sample; M is greater than or equal to 20 seconds; staining reagent A includes staining agent A10, adjusting salt C10, and water; staining agent A10 is neomethylene blue; the concentration of neomethylene blue in staining reagent A is 0.8 × 10⁻⁶. ‑4 g / ml ~ 8.0 × 10 ‑4 g / mL; adjusting salt C10 is used to provide a low osmotic pressure environment and a pH range of 7.0–8.0; stabilizing reagent B includes staining stabilizer B10, adjusting salt D10, and water; the staining stabilizer B10 in stabilizing reagent B includes aldehydes. A unique two-step reagent formulation and method are used to achieve rapid staining of reticulocytes in a step-by-step manner, achieving successful staining in just 20 seconds, and the staining effect can be maintained for a sufficiently long time, exceeding 120 minutes.

This invention relates to the field of immunoassay technology, specifically to a multicolor fluorescent diagnostic reagent for detecting PAP. The multicolor fluorescent diagnostic reagent includes Hurst dye, wheat germ lectin, fluorescein isothiocyanate, a counterstaining agent, an organic solubilizer, and a humectant. This invention incorporates multiple fluorescent dyes, enabling rapid and effective detection of glycoprotein signals in PAP patient specimens. Simultaneously, it clearly displays cell morphology in the specimens, allowing for timely exclusion of bacteria, fungi, and abnormal cells, thus helping doctors quickly and accurately determine the type of PAP in patients. Compared to the traditional PAS staining method, the multicolor fluorescent diagnostic reagent provided by this invention uses a one-step staining method, which is simple to operate, has a short detection time, and allows for rapid observation of test results, facilitating timely diagnosis and treatment for patients.

The invention provides a dyeing method for removing autofluorescence of a single cell sorting micro-fluidic chip and a chip without autofluorescence. The dyeing method comprises the following steps: completely soaking the single-cell sorting micro-fluidic chip in a dye solution for dyeing until the top layer of the chip is dyed into a non-transparent black top layer, thereby obtaining the single-cell sorting micro-fluidic chip without autofluorescence, the dye in the dye solution is black dye; the single cell sorting micro-fluidic chip at least comprises a photosensitive dry film serving as a runner layer. The staining method provided by the technical scheme of the invention can effectively solve the problem of autofluorescence of a single cell sorting micro-fluidic chip prepared from a traditional photosensitive dry film, so that when the stained chip is applied to fluorescence activated cell sorting, recognition of automatically screened fluorescent cells is facilitated, the operation difficulty of fluorescence activated cell sorting is reduced, and the working efficiency is improved. And accuracy and reliability are improved.

The invention relates to the field of biotechnology and dead / living cell detection, in particular to a method for rapidly determining the activity of biological membrane cells in immobilized fermentation. According to the method, a methylene blue and trisodium citrate synergistic chromosome system is optimized, a biofilm EPS matrix is synergistically penetrated by virtue of the dispersion effect of trisodium citrate and the reduction characteristic of methylene blue, and a spectrophotometric method is combined to detect the OD value of a staining solution instead of stained cells, so that the efficient quantitative analysis of the ratio of dead cells to living cells of the immobilized biofilm is realized. The method can complete detection within 200 s, is simple and convenient to operate and low in cost, does not need complex equipment, and is remarkably superior to a traditional fluorescent staining method.

The invention belongs to the technical field of antibody detection, and particularly relates to a multi-subtype antigen co-transfected cell immunofluorescence staining method and application. The method is established for the purpose of a CBA detection method, the basic principle is similar to that of an overexpression antigen cell immunofluorescence method for existing anti-GFAP-IgG antibody detection, and the difference is that three GFAP subtypes are adopted for cotransfection design, antigen subtypes are covered, and the problem of leak detection is avoided. According to the invention, the high-quality expression of the antigen is realized by self-constructing the plasmid based on the pCDH lentiviral vector. According to the invention, by optimizing the schemes of fixation, transparent preservation and the like after cell transfection culture, the dyeing performance is enhanced, and the preservation effect is improved.

To provide an improved verification method for verifying the performance of a fluorescence-based analysis system.SOLUTION: (a) negatively staining cells in a first portion of a standard sample comprising target cells expressing a target cell marker; (b) positively staining target cells in a second portion of the standard sample; (c) passing the first and second portions of the standard sample through an instrument to obtain a fluorescence measurement indicative of target cell concentration; (d) based on the fluorescence measurement obtained in (c), each diluted sample having a nominal concentration of target cells, each nominal cell concentration being greater than the concentration of target cells indicated by the fluorescence measurement of the negatively stained first portion in (a); Preparing a dilution series comprising a plurality of diluted samples, wherein the nominal concentration in each diluted sample is different from the nominal concentration in each of the remaining diluted samples; (e) passing the dilution series of (d) through the instrument and obtaining a fluorescence measurement; and (f) comparing the nominal cell concentration of (d) and the fluorescence measurement of (e) for each diluted sample to quantify the performance of the staining method in the instrument.SELECTED DRAWING: Figure 1

w The invention discloses a detection method for predicting efficacy and prognosis of immunotherapy for small cell lung cancer based on ATAD2 expression, and relates to the technical fields of molecular biology and cancer treatment. The method comprises the following steps: extracting a tumor tissue sample of a small cell lung cancer patient through surgical resection; Detecting the tumor tissue sample to obtain ATAD2 expression level, wherein the detection adopts an immunohistochemical staining method. According to the invention, the expression level of ATAD2 in tumor tissues of small cell lung cancer is quantitatively evaluated by immunohistochemical detection technology, and the molecular marker information is systematically correlated with the clinical and pathological characteristics of patients by combining Cox regression analysis and Logistic regression analysis, so that the comprehensive prediction of immunotherapy efficacy and patient survival time is realized.

The invention discloses an annona squamosa seed vigor grading detection method, and relates to the technical field of annona squamosa seed vigor detection, and the method comprises the following steps: 1, collecting a seed sample; 2, seed pretreatment; 3, measuring the constant weight of the seeds; 4, measuring the water absorption rate of the seeds; 5, modeling after detection, and detecting and recording multiple data of the annona squamosa seeds through a TTC staining method, a conductivity determination test and a germination test; and step 6, vigor grading standard formulation. According to the method, multiple annona squamosa seed groups are adopted for detection, and multiple data of annona squamosa seeds detected through the water absorption rate, the TTC dyeing method, the conductivity determination test and the germination test are recorded, so that the method can be more convenient and more accurate when modeling comparison is performed on the data of the annona squamosa seed groups. Therefore, when the method is used for detecting and grading the activity of the annona squamosa seeds, the method can be more accurate, and the accuracy of the method during detection is improved.

The invention discloses application of hyperoside to treatment of endometriosis, and aims to evaluate whether hyperoside has an anti-fibrosis effect when applied to a rat endometriosis model by taking fibrosis as a treatment target. The method comprises the following steps: constructing a rat endometriosis model through an autotransplantation method, dividing molded rats into a low-dose group, a medium-dose group, a high-dose group, a model group and a western medicine control group 4 weeks after successful modeling, and laparotomizing and taking out ectopic lesions after respectively taking medicines for 4 weeks. HE staining and masson staining methods are used for mutually proving the fibrosis degree, the expression conditions of fibrosis indexes a-SMA, Co-1, CTGF and Fn are detected in an immunohistochemical mode, and the anti-fibrosis effect of hyperoside is evaluated. As a result, hyperoside can prevent the development of fibrosis in endometriosis, and thus hyperoside can be used as a drug for treating or preventing fibrosis in endometriosis.

The invention discloses a coomassie brilliant blue staining solution as well as a preparation method and a staining method thereof. The coomassie brilliant blue staining solution comprises the following components: coomassie brilliant blue, sulfate, inorganic acid, an alcohol solvent, a polar amide solvent and water. The sulfate and the polar amide solvent are introduced into the coomassie brilliant blue staining solution; the sulfate can shorten the dyeing time and reduce the dyeing background; the polar amide solvent can improve the solubility of the coomassie brilliant blue staining solution, so that the staining efficiency and speed are improved, and the detection sensitivity is improved. The protein gel dyeing method is based on the improved coomassie brilliant blue dyeing solution, microwave heating is combined, the whole dyeing process of SDS-PAGE can be completed within 10 minutes, the steps of gel fixation, sensitization, decoloration and the like in a traditional dyeing procedure are omitted, the process steps are simplified, the dyeing time is saved, and the dyeing efficiency is improved. The method has the advantages of fast detection, high sensitivity and small background interference.

The invention belongs to the technical field of medical detection, and discloses a staining reagent applied to acid-fast staining and a staining method. The dyeing reagent comprises a primary dyeing agent, a decolorizing agent, a counterdyeing agent and an accelerating agent, wherein the accelerating agent is prepared from the following components: lauryl sodium sulfate, triton X-100, hexadecyl trimethyl ammonium bromide, a Tris-HCl buffer solution, ethylenediamine tetraacetic acid, sodium chloride, potassium chloride, protease K and beta-mercaptoethanol. According to the dyeing reagent, the dyeing accelerant is introduced into a traditional acid-fast dyeing system, the component capable of specifically damaging the lipid structure of the cell wall of the Mycobacterium is utilized, the hindering effect of the lipid structure on the dye in the dyeing process is reduced, the binding efficiency of the dye and thalli is remarkably improved, and therefore the positive detection rate of acid-fast dyeing is greatly increased.

The invention relates to the technical field of medical detection, in particular to an antibody combination for detecting spermatogenic cells, a kit and application of the antibody combination. The antibody combination comprises an antibody of a zinc finger transcription factor 4, an antibody of a conjugation complex protein 3 and an antibody of zona pellucida binding protein. Specific recognition and semi-quantitative analysis of spermatogenic cells of all levels in samples are realized through standardized pretreatment, full-automatic immunohistochemical staining and digital image analysis for different types of samples; the method breaks through the technical bottlenecks that the traditional HE staining or Wright-Giemsa staining method is strong in subjectivity, difficult to distinguish cells with similar forms and greatly influenced by cell states and staining effects, and provides an efficient detection means for accurate pathological typing, spermatogenic function evaluation and dynamic curative effect monitoring of male infertility.

The invention discloses a method for detecting sperm morphological anomaly through an improved Pasteur staining method, and relates to the technical field of morphological analysis, and the method comprises the following steps: carrying out staining treatment on a sperm smear by using the improved Pasteur staining method to obtain a training data set and a test data set; based on a U-Net network, designing a convolutional neural network VCA-Net applied to sperm morphological anomaly detection; based on the training data set and the test data set, performing model training and testing on the designed convolutional neural network VCA-Net to obtain a final convolutional neural network VCA-Net; and evaluating the final convolutional neural network VCA-Net by using the evaluation index to complete the anomaly detection of the sperm morphology by the improved Pasteur staining method. The problems that in the prior art, SMA automatic detection is difficult to achieve, and manual detection consumes time and labor and is large in workload and high in subjectivity under existing microscopic imaging are solved.

Proposed is a method for estimating the mass of microplastics by using fluorescent staining. The method may include staining microplastic samples by using a Nile red solution, and capturing a fluorescent image of each of the stained microplastic samples after classifying the stained microplastic samples by mass. The method may also include analyzing the captured fluorescent image to quantify a fluorescence intensity thereof to build a database (DB) about a correlation between mass and fluorescence intensity of microplastics. The method may further include estimating a mass value of an unknown microplastic sample by comparing a fluorescence intensity of a fluorescent image of the unknown microplastic sample obtained by performing the staining and capturing of the unknown microplastic sample with data stored in the database.

The invention discloses a micro-plastic organism in-vivo in-situ integrated analysis method based on phthalocyanine blue labeling and Raman imaging, and belongs to environmental toxicology and analytical chemistry technologies. The method comprises the following steps: firstly, exposing organisms to be researched and analyzed in phthalein blue dyed micro-plastic particles; then obtaining exposed biological tissues, preparing paraffin sections and staining the tissues; positioning a biological tissue area under an optical microscope, acquiring a spectrum of the area by adopting a Raman spectrum point scanning mode, and confirming a target area by identifying characteristic Raman peaks of phthalocyanine blue and micro-plastic; raman surface scanning is carried out on a target area, intensity information of characteristic Raman peaks of phthalocyanine blue or micro-plastic is obtained through a peak clamping method, a spatial distribution image of corresponding components in biological tissue is generated, and quantification of the micro-plastic can be achieved. According to the method, the problem that the marking stability of a traditional plastic dyeing method is difficult to be compatible with a standard slide making process is solved, in-situ association of micro-plastic distribution and tissue lesion can be realized, and the method is suitable for accurate detection of submicron micro-plastics.

The present application relates to the technical field of cell staining, and particularly relates to a vaginal secretion staining solution, a preparation method and a staining method thereof.The vaginal secretion staining solution provided by the present application uses basic fuchsin and methylene blue as staining agents, can stain epithelial cells and leukocytes, and cannot stain trichomonas and mold, so that leukocytes and trichomonas can be obviously distinguished through color difference, and personnel misjudgment caused by the similarity of leukocytes and trichomonas is reduced.The vaginal secretion staining solution provided by the present application has simple operation steps, short cycle and fast staining.The present application further provides a preparation method and a staining method of the vaginal secretion staining solution.

The invention relates to the technical field of biology, and particularly discloses a staining method and application of an immunohistochemistry multi-staining combined reagent, the combined reagent comprises a first antibody combination and a second antibody combination, the first antibody combination comprises a myoepithelium specific marker Calponin and p16 with myoepithelium expression characteristics and a cell cycle regulation function; and the second antibody combination comprises an epithelial marker E-cadherin. According to the invention, three markers of Calponin, p16 and E-cadherin are subjected to immunohistochemical multi-staining combination, so that whether muscle epithelial cells exist or not and the attributes of gland epithelial cells can be synchronously displayed on the same tissue slice at the same site; mutual-verification and complementary morphological and molecular basis is provided for indirectly prompting whether the basilar membrane is damaged or not through the integrity of the muscular epithelial layer, and the detection rate of the micro-invasive cancer is increased. Meanwhile, spatial distribution information of the three types of key markers can be obtained through one dyeing process, tissue samples are saved, and the method is suitable for accurate diagnosis of biopsy specimens.

The invention discloses a method for separating and identifying a peripheral blood circulating tumor cell cluster based on a density gradient centrifugation method. The method comprises the following steps: (1) molding mouse tumor cells; (2) taking blood from the heart of the modeling mouse, and diluting; (3) separating by adopting a density gradient centrifugation method, and taking a lymphocyte layer and a red blood cell layer after layering; (4) centrifugally washing to obtain lymphocyte precipitate and erythrocyte precipitate; (5) adding a red blood cell lysis solution into the red blood cell precipitate, centrifuging to obtain a cell precipitate 1, adding a serum-free culture medium, and centrifugally washing to obtain a cell precipitate 2; (6) resuspending and culturing the lymphocyte precipitate and the cell precipitate 2 by using a culture medium; and (7) replacing the culture medium, wherein the remaining adherent cells are cell populations containing peripheral blood circulating tumor cells and circulating tumor cell clusters. The CTC cluster can be successfully separated, the condition of separation failure caused by loss of the CTC cluster is reduced, the CTC cluster is identified by using an immunofluorescence staining method, and the method is simple and high in operability.

The invention belongs to the technical field of dairy product detection, and particularly relates to a method for detecting the proportion and activity of each whey protein in milk and dairy products through a fluorescent gel electrophoresis technology. Carrying out lauryl sodium sulfate-polyacrylamide gel electrophoretic analysis on the milk and dairy products marked by the protein fluorescent dye, firstly adding concentration gel for concentration, compressing the protein dispersed in a sample loading hole into a narrow sample band, then adding separation gel for separation, and according to the difference of molecular weights of different whey proteins and casein, determining the content of the whey protein and the casein. According to the method, whey proteins are separated from each other, migration rates in gel are different, effective separation is realized, finally, an electrophoretogram is obtained through a gel imaging system, a gray value of the electrophoretogram is subjected to quantitative processing by adopting gel image analysis software, and the proportion and activity of each whey protein are obtained through calculation. Compared with a traditional Coomassie brilliant blue dyeing method, the method is high in marking speed, does not need time-consuming dyeing and decolorizing steps, and is high in practicability.

The invention relates to the technical field of biological tissue staining, in particular to a staining method and a staining reagent for staining a pituitary gland of a rat, and the staining method adopts a multi-dye synergistic combination of PAS staining, hematoxylin staining, fast green staining and orange G staining to stain a pituitary gland tissue section of the rat. According to the dyeing method disclosed by the invention, the rat pituitary tissue section is dyed through the synergistic combination of multiple dyes, and the core defects of fuzzy details, unclear boundary of suspected cells, light dyeing and low identification degree of traditional dyeing particles are radically solved through the precise synergy and functional complementation of the dyes.

The application discloses a vaginal micro-ecological detection staining method, mainly comprising the following steps: 1) taking female genital tract secretion and diluting the same by a diluent to form a suspension; 2) coating the suspension on a glass slide; 3) drying and fixing the suspension on the glass slide; 4) immersing the glass slide in a eosin dye solution for staining, and then taking out the glass slide and washing the same by flowing water; 5) immersing the glass slide in a methylene blue dye solution for staining, and then taking out the glass slide and washing the same by flowing water; 6) performing gram staining on the glass slide; and 7) observing by using a microscope. The staining method can not only clearly show the trophozoite shape and flagellum shape of Trichomonas vaginalis, but also accurately distinguish leukocytes and trichomonads, is favorable to improving the accuracy of trichomonas vaginalis detection, and is favorable to comprehensively and accurately detecting the vaginal micro-ecology.

The present application relates to the technical field of in vitro detection, in particular to a rapid sperm morphology staining reagent and a staining method thereof; the rapid sperm morphology staining reagent of the present application can make semen spread rapidly and uniformly after 5 minutes by using bromelain and L-cysteine in combination. The staining reagent of the present application greatly shortens the pretreatment time of the sample and improves the detection efficiency.

This invention discloses a lignin-degrading microorganism, its isolation method, and its application, relating to environmental microbiology technology and the field of biodegradation. The isolation method includes: enriching the sample to be isolated using alkaline lignin as the sole carbon and energy source; then using morphological recognition-based single-cell sorting technology and resazurite staining to screen for active core strains that grow well in a single lignin culture medium and can dissolve lignin precipitates; and finally, sequencing identification to obtain a pure microbial strain that degrades lignin. This isolation method can effectively isolate lignin-degrading microorganisms, is simple and efficient, and is suitable for large-scale applications. Furthermore, this invention identifies [specific microorganisms] based on this isolation method. Phytobacter ursingii It is an active core strain for degrading lignin, with outstanding lignin degradation efficiency, providing an important strain resource for developing microbial-based biological treatment technology for lignin waste.

The application provides a method for quantitatively analyzing a greasy fur. Based on previous research, the inventors found that the essence of the greasy fur is that the tongue surface bacteria form a layer of biofilm between the tongue filiform papillae, and firmly adhere to the filiform papillae. Based on the detection method of the biofilm, the tongue fur samples are respectively taken from the greasy fur and the normal tongue fur, cultured in vitro, and the number of biofilm formation is detected, and it is found that the bacterial biofilm of the greasy fur is much higher than that of the normal tongue fur. Therefore, by detecting the biofilm formation of the tongue fur microorganism, or scraping part of the tongue fur smear, and detecting the polysaccharide components representing the sticky properties of the biofilm in the tongue fur smear by the fluorescent dye method and the AB / PAS mucopolysaccharide staining method, the strength of the greasy fur can be quantitatively detected in a high-throughput and rapid manner.