37 results about "Specific staining" patented technology

The invention relates to the technical field of chemiluminescence immunoassay, and discloses a horse radish peroxidase-labeled secondary antibody compound and a preparation method and application thereof, the horse radish peroxidase-labeled secondary antibody compound comprises a chitosan compound skeleton, and horse radish peroxidase and a secondary antibody carried on the chitosan compound skeleton; the chitosan compound skeleton is a macromolecular skeleton compounded by a chitosan spherical cross-linking body and dopamine. Through optimization and improvement of a polymer skeleton, the problems of large steric hindrance, low sensitivity and non-specific staining of a traditional polymer can be solved, and further improvement of the sensitivity of a DAB staining method for immunoassay detection is facilitated; the adopted chitosan compound skeleton has abundant amino groups, can bear more labeled enzymes and secondary antibodies, reduces steric hindrance, increases the penetrating power to a cell nuclear membrane, and improves the detection sensitivity.

This invention proposes a sperm staining kit based on a sequential staining strategy of tail pre-staining – rapid differentiation – enhanced nuclear staining. A gentle fixation with 4% paraformaldehyde and acetate buffer preserves the complete nucleic acid of the sperm head and the protein structure of the tail. The addition of PVP K30 and Tween-20 effectively inhibits non-specific adsorption and reduces staining background interference. Brilliant Green SF combined with sodium tartrate is used for specific staining of the sperm tail, followed by rapid differentiation to wash away non-specific staining. Nuclear staining is performed using toluidine blue O combined with sodium chloride and sodium tetraborate hydrate, specifically binding to DNA and enhancing color development stability, resulting in a blue sperm head and a light green tail with clear color contrast. This invention produces a clean staining background, accurately distinguishing the morphology of the sperm head, acrosome, and tail, significantly improving the accuracy of microscopic identification of abnormal sperm such as large-headed, small-headed, curled-tailed, broken-tailed, and short-tailed sperm. It is suitable for detecting sperm morphology and abnormality rates in breeding males.

The invention provides a multiplex fluorescence immunohistochemical detection kit for rare gastrointestinal tumors and application of the multiplex fluorescence immunohistochemical detection kit, and belongs to the technical field of multiplex immunohistochemistry. The invention provides a multiplex fluorescence immunohistochemical detection reagent combination, which comprises a monoclonal antibody designed for protein markers and a fluorescent dye, and can be applied to multiplex fluorescence immunohistochemical detection for detecting the protein markers such as PD-L1, PD-1, CTLA-4, LAG-3, TIM-3 and TIGIT. The invention further provides a kit capable of being used for multiplex fluorescence immunohistochemical detection, staining detection of six protein markers on a single section can be achieved, and multiplex fluorescence staining of disease samples, especially complex-structure gastrointestinal tract rare tumor samples is achieved by matching six protein targets according to a specific staining sequence and fluorescence channels. And false positive caused by staining crosstalk of each channel is prevented.

Proposed are a sessile droplet biosensor and an extracellular vesicle detection method using same, wherein the sessile droplet biosensor can easily and conveniently perform superbright staining of proteins or lipids in extracellular vesicles through a non-specific staining material, such as CFSE, without a complicated signal generation process and can concentrate extracellular vesicles to a high concentration at the edges of sessile droplets by internal flowing induced by non-uniform evaporation in the sessile droplets, thereby detecting extracellular vesicles with high sensitivity. Moreover, the extracellular vesicle detection method using the sessile droplet biosensor can be utilized for standard setting technology for various diseases, such as cancer diagnosis standard setting technology, by the analysis of extracellular vesicle staining signals, or an information providing method for the analysis of extracellular vesicle staining signals can be utilized for early diagnosis of various diseases such as cancer, evaluation of prognosis for treatment, and screening for carcinoma.

The present invention discloses a method for testing the adhesion of jellyfish nematocysts and its application. The method comprises the following steps: isolating nematocysts: subjecting a jellyfish suspension to density gradient centrifugation using Percoll centrifuge to obtain nematocysts; preparing a coating: applying different coating materials to the center of a culture dish to form a smooth coating; adhering nematocysts: dropping the separated nematocysts onto the center of the coating and covering it with a coverslip for a predetermined period of time; staining: removing the coverslip, adding acridine orange as a dye, covering it with a coverslip to allow the dye to spread, and staining for a predetermined period of time; and image processing: photographing the sample using an inverted fluorescence microscope and processing the image using image processing software to obtain the fluorescently stained area. The method uses acridine orange for staining, resulting in specific staining, clear images, and easy identification of nematocysts. The results can be digitized. This method is rapid, simple to use, and highly operational, making it easy to screen for jellyfish sting protective coatings.

The invention relates to a 177Lu-DOTATATE peptide receptor radionuclide curative effect prediction model and application thereof, the model is used for performing multicolor immunofluorescent staining on tissue slices of a neuroendocrine tumor (NET) patient, and the model at least comprises specific staining antibodies of cytokeratin (CK +) cells, CD8 cells and TIM-3 molecules; then carrying out image acquisition and analysis, and identifying two positive cells, namely CK < + > and CD8 < + > TIM-3 < + >; calculating an under-curve area difference AUC (CK) (CD8 + TIM-3 +) between an observation curve of a nearest neighbor distance accumulation function (G function) between CK + and CD8 + TIM-3 + and a Poisson distribution curve according to an R language software package, determining a response scoring formula and threshold division based on single-factor logistic regression of PRRT response degree of case data, and substituting the difference into the response scoring formula, and the response score can be used for curative effect prediction.

The utility model discloses tissue immunofluorescence bleaching and dyeing equipment, which relates to the technical field of immunofluorescence and comprises a bottom plate, a cover plate is arranged on the upper side of the bottom plate, a groove is arranged on the upper side surface of the cover plate, the bottom end of the cover plate penetrates into the groove and is provided with a drain hole, and a water blocking plug is arranged on the upper side surface of the bottom plate and penetrates through the drain hole. According to the utility model, waste of tissues during slicing is avoided, a plurality of samples can be treated at the same time, the tissues can be dyed more fully and thoroughly, and non-specific dyeing can be reduced.

The invention discloses a multiple immunofluorescence color development kit and application thereof, the multiple immunofluorescence color development kit comprises a fluorescent material, the structure of the fluorescent material is as shown in formula (I), the fluorescent material in the multiple immunofluorescence color development kit has a weak fluorescence signal in the absence of HRP enzyme catalysis, and does not generate non-specific adsorption staining on tissue slices, so that the problem of non-specific staining in the use process of existing immunofluorescence histochemistry can be solved.

The invention discloses a multiplex fluorescence immunohistochemical detection kit for rare lymphoma and application of the multiplex fluorescence immunohistochemical detection kit. The multiplex fluorescence immunohistochemical detection kit for rare lymphoma comprises a primary antibody reagent and a fluorescence staining reagent, the primary antibody reagent is prepared from a rabbit anti-human CD15 monoclonal antibody, a rabbit anti-human CD30 monoclonal antibody, a KI-67 monoclonal antibody, a rabbit anti-human CD79b monoclonal antibody, a rabbit anti-human CD3 monoclonal antibody and a rabbit anti-human CD20 monoclonal antibody. When the multiplex fluorescence immunohistochemical detection kit for the rare lymphoma is used, a false positive result caused by channel spectrum crosstalk is avoided to the greatest extent through a specific staining sequence of each target protein and fluorescein matching, and the purpose of detecting the rare lymphoma, especially the rare type of lymphoma, is achieved. The kit can be used for synchronous detection of lymphoma, such as hodgkin lymphoma, mantle cell lymphoma and marginal region lymphoma, and provides a reliable basis for lymphoma immune targeted therapy.

The application provides a prognosis risk stratification prediction method for lymph node tumor residue, and belongs to the technical field of risk prediction, and comprises the following steps: acquiring lymph node HE staining sections and immunohistochemical labeling sections, automatically identifying a tumor region of interest by using a U-Net++ network, removing background noise and non-specific staining interference based on a self-adaptive threshold segmentation algorithm, extracting information in four dimensions, and forming a standardized extracted information set; fusing and mapping the extracted information set into a 256-dimensional feature vector set, inputting each vector into a dynamic integrated prediction model, and outputting stratified differential expression of each vector and stratified comprehensive expression of the vector set; determining a risk level based on quantitative threshold values of the stratified differential expression and comparative analysis of risk probabilities of the stratified comprehensive expression, and constructing a multi-dimensional dynamic weight decision model in combination with patient self-state indexes to formulate a personalized scheme. Through multi-modal feature extraction, dynamic integrated modeling and personalized decision-making, precise risk stratification and treatment scheme customization are realized.

This invention discloses a method for detecting Candida albicans based on compound fluorescent staining and deep learning, belonging to the field of microbial detection technology. The method first synthesizes a novel NHC-triazolium salt fluorescent molecule, which is then compounded with calcium fluorescent white and DAPI to form a specific staining solution. After treating the sample with this staining solution, multi-channel fluorescence imaging is performed and images are acquired. Simultaneously, ITS sequencing is performed on the samples. Based on the sequencing results, the images are classified and bacterial cells are manually cropped to construct a labeled dataset. A convolutional neural network model is trained using this dataset to learn to recognize the unique "multi-fluorescent fingerprint" presented by Candida albicans stained with the compound staining solution. In application, the same staining and imaging are performed on the test samples, and the trained model automatically analyzes the images to achieve detection. This invention significantly improves the specificity of detection through the synergy of chemical staining and artificial intelligence. Experiments show that compared with traditional single staining or unstained methods, this method improves the specificity of Candida albicans recognition by 37% and 52%, respectively, and has the advantages of being fast, accurate, and highly automated.

The invention relates to the technical field of dyeing characterization of surface functional layers of composite materials, in particular to a hydrophilic functional layer specific coloring agent and a preparation method and application thereof, and the hydrophilic functional layer specific coloring agent is composed of 0.01%-0.5% by mass of xanthene fluorescent dye, 0.1%-5% by mass of hydrophilic polymer additive and the balance of water. Wherein the xanthene fluorescent dye is 9-(2-carboxyl phenyl)-3, 6-bis (diethylamino) xanthyleione chloride, and the structural formula of the xanthene fluorescent dye is shown in the description. After a hydrophilic film is dyed by the specific dyeing agent for the hydrophilic functional layer, an extremely high color contrast can be presented between the hydrophilic functional layer and a base material layer, and the background is clean and clear. For the defects such as uneven coating, microcracks and pinholes which cannot be recognized by conventional visual observation and are in the order of nanometer to micrometer, the coloring agent can clearly expose the defects through the difference of local color depth or fluorescence intensity, and visual precision diagnosis of the quality of the hydrophilic functional layer is achieved.

This invention discloses a large-section kit for pathological prostate tissue, comprising an embedding cassette, an embedding mold, and a sample holder disposed within a storage box. The embedding cassette includes an embedding cassette body, a first partition, a first elastic clamping device, an embedding cassette lid, and a mounting ring. The bottom plate of the embedding cassette body, the embedding cassette lid, and the first partition are all plate structures with mesh openings. The pathologically obtained prostate tissue is placed between the first partition and the bottom plate of the embedding cassette body, and the elastic clamping device is disposed between the first partition and the embedding cassette lid. The embedding mold includes an embedding mold body, a second partition, and a second elastic clamping device. This invention can effectively improve the integrity and consistency of slide preparation, reduce specimen loss, and ensure the homogeneity of continuous step slides. Based on this, various specific staining methods can be performed, improving the reliability of pathological clinical diagnostic assessment and the objectivity of biological information diversity analysis in basic research, thereby reflecting the reliability of the gold standard for pathological testing.

The application discloses a kind of portable microplastics rapid detection method and device based on fluorescence, including sampling module, pre-treatment module, fluorescence detection module and control module;Sampling module is connected with pre-treatment module, and sampling module is used for the collection of microplastics in water body;Pre-treatment module is used to remove redundant impurities, retain microplastics, and specifically dye it;Fluorescence detection module is connected with pre-treatment module, and the application relates to the technical field of water quality detection.This kind of portable microplastics rapid detection method and device based on fluorescence, the portable microplastics rapid detection device, high degree of automation, can complete the qualitative and quantitative analysis task of microplastics quickly and accurately on site, avoid the time and cost overhead of sample return laboratory analysis, save time and effort, not only can effectively improve the efficiency of sample microplastic detection, but also greatly reduce the cost of sample microplastic detection.

This invention discloses a method and system for in vitro assessment of renal biopsy tissue quality based on virtual special staining, belonging to the field of biomedical image processing technology. The method includes: acquiring and scanning undewaxed mounted sections to be assessed to obtain unstained whole-section images; inputting the unstained whole-section images into a pre-trained hybrid contrast virtual staining generation model to generate virtual special staining images with specific staining textures; the hybrid contrast virtual staining generation model is obtained through optimization training based on a total loss function, which includes adversarial loss, L1 loss, and PatchNCE block noise contrast estimation loss to maximize the mutual information between the input and generated images at corresponding spatial locations; inputting the virtual special staining images into a target detection network to identify glomerular structures and count the total number of glomeruli; and outputting the suitability assessment result of the undewaxed mounted sections based on the comparison result between the total number of glomeruli and a preset threshold. By scanning undewaxed mounted sections, the traditional dewaxing and rehydration steps are eliminated, improving efficiency; and the virtual silver staining technology significantly enhances the ability to identify glomeruli.

The invention discloses a full-spectrum flow type monitoring method for peripheral blood circulating tumor plasma cells, and relates to the technical field of biomedical detections.The monitoring scheme comprises the specific steps that S100, sample collection and preparation are conducted, specifically, 5-10 ml of peripheral blood is collected from elbow veins of a patient according to sterile operation and put into a vacuum tube containing an anticoagulant, and then the peripheral blood is collected; and transporting at low temperature, and storing at 4 DEG C for a short time or at-80 DEG C for a long time according to conditions. According to the present invention, with the optimized antibody combination, the introduction of the high-specificity tumor-associated antigen such as BCMA, and the combination of the multi-color staining and the spectrum deconvolution technology of the full spectrum flow cytometry, the dye overlapping is avoided, and the detection ability and the resolution of the multi-labeling are improved, such that the extremely low-content CTPCs can be reliably identified, and the detection sensitivity is high; the interference of non-specific staining is reduced, the generation of false positive and false negative results is reduced, and an accurate basis can be provided for early diagnosis of multiple myeloma and other plasma cell tumors.

A semi-supervised tissue slice image virtual staining method belongs to the technical field of image processing algorithms, and has the specific scheme as follows: specific staining of different tissues in the same sample is realized by marking pixel-level tissue types in an original image and a stained image and introducing supervision information in a training process of a generative network. The supervision mechanism based on the tissue type can significantly improve the accuracy of the dyeing result and the tissue difference, so that the generated dyeing image is closer to the actual dyeing effect, and the high requirement on the dyeing specificity in pathological analysis is met. In order to reduce the cost of data annotation, the invention designs a semi-supervised training mode. According to the method, the dyeing neural network can be effectively trained on the premise that only part of the images in the data set need to be organized and labeled, and the tedious process of full data set labeling is avoided. The innovation reduces the time and labor cost of data preparation, and provides convenience for popularization in practical application.

The application discloses an auxiliary lymphoma immunohistochemical positive cell positioning analysis device and method, and relates to the technical field of positive cell positioning.The application comprises an image acquisition and preprocessing module, a positive area preliminary screening module, a cell morphology analysis module, a three-dimensional coordinate positioning module and a report generation and visualization interaction module, is used for accessing a digital slice scanner or a digital pathology image management system, reading an immunohistochemical slice image in a WSI format, and being internally provided with a convolutional neural network model based on deep learning; the model is trained and optimized through a large number of annotated lymphoma immunohistochemical slice images, and can automatically identify suspicious areas with positive cell characteristics in the image.The double-layer analysis mechanism combining the deep learning model and the multi-dimensional screening of morphological parameters can significantly reduce the interference of non-specific staining and background impurities, and effectively reduce the missed detection rate and the false detection rate, compared with the traditional manual film reading or single threshold judgment method.

This invention relates to the field of image analysis technology, specifically to a method for assessing the distribution of cardiac mesothelial cells based on immunofluorescence images. The method includes: acquiring multiple immunofluorescence images of cardiac mesothelial cells at different antibody concentrations within the assessment group; determining initial cardiac mesothelial cell regions based on the staining status of the isotype control group and the differences in staining depth among multiple staining regions in the immunofluorescence images; analyzing non-specific staining based on changes in the staining area and irregular cell boundaries of the initial cardiac mesothelial cell regions, and adjusting the antibody concentration accordingly; determining multiple target cardiac mesothelial cell regions, and obtaining the location information of the cardiac mesothelial cells based on the degree of overlap and similarity between these regions; and analyzing the spacing distribution of cardiac mesothelial cells and their distribution in the immunofluorescence images based on the location information to obtain the uniformity of the cardiac mesothelial cell distribution.

The purpose of the present invention is to provide a method for evaluating the clogging of a filtration membrane with a protein-containing solution, including the steps of:a) passing a protein-containing solution through a filtration membrane,b) after the step a), obtaining a filtration membrane cross-section from the filtration membrane,c) treating the protein-containing solution before the step a), the filtration membrane before the step b), or the filtration membrane cross-section after the step b) with at least one stain specific to a protein aggregate, andd) confirming the presence of the protein aggregate in the filtration membrane cross-section.

The invention relates to the technical field of image analysis, in particular to a cardiac mesothelial cell distribution assessment method based on immunofluorescence images, which comprises the following steps: acquiring a plurality of immunofluorescence images corresponding to cardiac mesothelial cells under different antibody concentrations in a group to be assessed; determining an initial heart mesothelial cell region according to the dyeing condition of the same type control group and the dyeing depth difference of a plurality of dyeing regions in the immunofluorescence image; analyzing the non-specific staining condition according to the staining area change and the cell boundary irregularity condition of the initial heart mesothelial cell region, and adjusting the antibody concentration; determining a plurality of target cardiac mesothelial cell regions, and obtaining the position information of cardiac mesothelial cells according to the overlapping similarity among the plurality of target cardiac mesothelial cell regions; and according to the position information of the heart mesothelial cells, analyzing the spacing distribution of the heart mesothelial cells and the distribution of the heart mesothelial cells in the immunofluorescence image to obtain the distribution uniformity of the heart mesothelial cells.

The invention discloses application of an MRPS10 inhibitor in diagnosis and treatment of pancreatic ductal adenocarcinoma, and relates to the technical field of biological science. The preparation method of the MRPS10 inhibitor comprises experimental design verification and preparation of the MRPS10 inhibitor, and the experimental design verification comprises the following steps: S1, immunohistochemical staining: performing labeling, dewaxing, hydration, antigen repair, endogenous peroxidase blocking, non-specific staining blocking, primary antibody incubation, secondary antibody incubation, color development and sheet sealing on a paraffin tissue section; s2, cell culture: cell resuscitation, liquid change, passage and cryopreservation operations are included. According to the invention, the effect of MRPS10 in PDAC is systematically analyzed, and the potential of MRPS10 as a potential therapeutic target is disclosed; in combination with bioinformatics analysis and various experimental methods, the function of the MRPS10 in the PDAC is comprehensively verified.

The present invention provides a histological staining method for plants of the genus Phyllostachys and a method for distinguishing phloem cells, and belongs to the technical field of tissue sections. The present invention provides a histological staining method for plants of the genus Phyllostachys, which uses antibodies generated by trimethylated histones for histological staining, and uses tissue sections as samples. The antibodies are expressed and undergo color development in cells other than the phloem, but are not expressed in the phloem, resulting in no coloring. Therefore, the method of the present invention can perform specific staining on different tissue cells, has high sensitivity, and is low in cost. An ordinary electron microscope can be used to determine whether the cell is a phloem cell based on whether the cell wall and / or cell nucleus are colored, and because the cell wall of the phloem is not colored, the cell nucleus is clearly visible. The histological staining method of the present invention is particularly suitable for detecting the morphological characteristics of the cell nuclei of early phloem cells.

The invention provides a histological staining method of phyllostachys plants and a method for distinguishing phloem cells, and belongs to the technical field of tissue slices. The invention provides a histological staining method of a phyllostachys plant, which is characterized in that histological staining is carried out by using an antibody generated by trimethylated histone, a tissue slice is taken as a sample, and the antibody is expressed in other cells except the phloem and is subjected to chromogenic reaction, but is not expressed in the phloem and is free from any coloration. Therefore, the method disclosed by the invention can be used for specifically staining different tissue cells, is high in sensitivity and low in cost, can be used for judging whether the cells are phloem cells or not by utilizing a common electron microscope according to whether cell walls and / or cell nucleuses are colored or not, and the cell nucleuses are clearly visible due to the fact that the cell walls of the phloem are not colored. The histological staining method disclosed by the invention is particularly suitable for detecting morphological characteristics of cell nucleuses of early-stage phloem cells.

The invention provides a paraffin section elastic fiber overdyeing basic fuchsin composite dyeing kit and a dyeing method thereof, and belongs to the technical field of pathology, a dyeing reagent in the dyeing kit is prepared from the following raw materials in percentage by weight: 4% of basic fuchsin, 50% of AR pure methanol, 0.3% of thymol, 0.6% of chloral hydrate, 1.5% of phenol and 43.6% of pure water. The dyeing step is simplified, the dyeing time is shortened, the experiment cost is reduced, the pathological diagnosis efficiency is improved, the dyed elastic fiber is specifically red, the display is clear, the background is clean, the stability and repeatability of the dyeing effect are improved, the dyeing error is reduced, the dyeing specificity is improved, non-specific dyeing is reduced, and observation and diagnosis are facilitated.

The application discloses a method for preparing a cell block by using mucin emulsifier and application thereof, and the preparation method comprises the following steps: centrifuging a non-gynecological sample to obtain a first separation liquid, adding the first separation liquid into a formaldehyde-alcohol solution to centrifuge to obtain a second separation liquid, and adding the second separation liquid into mucin emulsifier to centrifuge to obtain a cell block. The preparation method is simple, the manufacturing cost is low, the cell block has high enrichment efficiency, cell loss is small, and the mucin emulsifier does not affect cell activity. The cell block prepared by using the mucin emulsifier has the following advantages: the appearance of the section of the cell block is complete, the section has a regular circular structure, the porosity is moderate, and the thickness is uniform; the application effect of the section in two kinds of staining is good: the hematoxylin-eosin staining has a clear background and clear contrast, cells are uniformly distributed in a circular area, cell morphology presents a stereoscopic effect, and the excellent rate of the stained section is 98.94%; the immunohistochemical staining has a clean background, and structures and cells are specifically dyed, and the excellent rate of the stained section is 99.30%, so that the cell block has a wide application prospect.

The present application belongs to the technical field of biological detection, and particularly relates to a natural active collagen rapid identification kit, an identification method and application thereof. The rapid identification method and the kit are used for specific staining of active collagen with an intact triple helix structure by using a specific staining agent, and a visual dye-collagen complex precipitate is generated. By observing the color, state and amount of the precipitate, the active collagen with the intact triple helix structure can be rapidly qualitatively detected. Compared with the existing detection method, the rapid identification method simplifies the process, shortens the original 1-hour detection to 15 minutes, replaces picric acid with acid and alcohol double reagents, and makes the identification more intuitive, clear and accurate. The naked eye identification does not need to rely on instruments and equipment, and is more conducive to on-site rapid identification.

The staining condition evaluation method comprises: a step for imparting a first primary antibody to a test specimen under a first staining condition (step S11); a step (step S13) for capturing an image of the test specimen to acquire a first image; a step (step S15) for imparting a second primary antibody to the test specimen under a second staining condition; a step (step S17) for capturing an image of the test specimen and acquiring a second image; and a step (step S18) for determining whether or not the first staining condition and the second staining condition are appropriate on the basis of the detection results of the stained cells in the first image and the second image. In step S18, when the cells on the test specimen are specifically stained in the first image, the cells on the test specimen different from the first image are specifically stained in the second image, and the cells on the test specimen stained in the first image are not stained in the second image, it is determined that the first staining condition and the second staining condition are appropriate.

Described herein is an imaging-based method for identifying RNA molecules in a sample, which is able to determine both the location and sequences of the RNA molecules. The method comprises: forming randomized barcode attached to the RNA molecules in the sample by sequential stepwise addition of permanent nucleic acid adapters randomly selected from a pool; acquiring imaging signals from the sequence-specific staining of the newly added permanent nucleic acid adapters after each addition of a permanent nucleic acid adapter, which links the location information of the RNA molecules with the barcoding; sequencing the barcoded RNA molecules, which links the sequences of the RNA molecules with the barcoding; and matching the location of the RNA molecules to the sequences thereof by matching the barcoding. Also described are kits for performing the method.

This invention relates to the field of microbial detection technology, specifically to a method and apparatus for simultaneous culture and staining of fungi. This invention creatively proposes a culture-integrated staining scheme, premixing a specific staining agent A at an optimal concentration in the culture medium. In use, only sample inoculation and culture are required, followed by direct in-situ microscopic observation, eliminating the need for any subsequent picking, smearing, and staining steps. This method significantly simplifies the operational process, improves detection efficiency and biosafety, and, due to its low cost and effectiveness against a variety of common pathogenic fungi, has promising prospects for clinical application and widespread adoption.