5 results about "Erythrocyte lysate" patented technology

The invention discloses a lymphatic endothelial cell cyst fluid extraction method for lymphatic deformity, and belongs to the technical field of biological cell extraction. Comprising the following steps: S1, sample collection: collecting 5-10 mL of cystic fluid of a patient with macrocystic lymphangial malformation, and placing the collected cystic fluid in a sterile container; s2, primary centrifugation: centrifuging at a low speed of 1000rpm for 10 minutes, and retaining cell precipitates; s3, red blood cell lysis: adding a red blood cell lysis solution, placing at 4 DEG C for 3-5 minutes, centrifuging, and washing and purifying with PBS (Phosphate Buffer Solution); s4, cell culture: resuspending the cells in a culture medium containing an endothelial growth factor replenishing liquid, culturing at 37 DEG C in 5% CO2, and changing the liquid for 24 hours to remove non-adherent cells; and S5, cell identification: carrying out immunofluorescence staining identification 48-72 hours after cell adherence. The method is simple, convenient and efficient to operate, does not need complex digestion, can enrich high-purity and high-activity lymphatic endothelial cells, provides a stable cell model for related pathogenesis research and drug screening, and has important scientific research and clinical value.

The invention relates to the technical field of biomedicine, in particular to a trace blood DNA (deoxyribonucleic acid) extraction method which comprises the following steps: S1, mixing a blood sample and lysate according to a ratio of 1: 3, uniformly mixing by vortex, and centrifuging; s2, protease K is added, vortex mixing is carried out, incubation is carried out at 55 DEG C for 15 min, centrifugation is carried out for 30 s, and 500 microliters of a binding solution is added; and S3, completely transferring the liquid into a purification column with a collection tube, and standing for 2 minutes. The extraction method of the trace blood DNA comprises the following steps: denaturating protein through SDS (Sodium Dodecyl Sulfate), and separating nucleic acid; the binding liquid adopts high salt and low PH, so that the long fragment nucleic acid can be more easily bound on the adsorption column, guanidine hydrochloride can enhance the splitting effect, and the nucleic acid in the blood sample can be fully released due to the dual splitting effect; and on the other hand, the key point is that the step of removing red blood cells by red blood cell lysate is avoided, but the whole blood is directly lysed, so that the loss of white blood cells is reduced, the operation steps are simplified, and the method is suitable for extracting fresh and frozen anticoagulant blood samples.

The application provides a preparation method and application of a liver tumor tissue single-cell suspension, and the preparation method comprises the following steps: step 1, pretreating liver tumor tissue; step 2, step-by-step enzymolysis; the pretreated liver tumor tissue in step 1 is subjected to step-by-step enzymolysis by using an enzyme solution, and the enzyme solution in each step is collected after filtration to obtain a cell suspension; step 3, gradient centrifugation; the cell suspension is subjected to gradient centrifugation by using a cell separation solution to separate immune cells and liver parenchymal cells; step 4, red blood cell lysis; the immune cells obtained in step 3 are subjected to red blood cell lysis by using a red blood cell lysis solution, and centrifugal washing is carried out to obtain an immune cell precipitate; step 5, collecting the liver parenchymal cells in step 3 and the immune cell precipitate in step 4, washing the liver parenchymal cells and the immune cell precipitate by using a cell washing solution, centrifuging to obtain a cell precipitate, resuspending the cell precipitate by using a cell resuspension solution, and filtering to obtain the liver tumor tissue single-cell suspension.

This invention belongs to the field of biomedical technology, specifically relating to a method and kit for improving the sensitivity and specificity of circulating tumor cell (CTC) detection. The method first treats anticoagulated blood with erythrocyte lysis buffer, centrifuges to obtain a cell suspension (including leukocytes and circulating tumor cells); then, a radiosensitizer is added and the suspension is irradiated; next, single-cell gel electrophoresis is used to detect the degree of DNA damage in the test cells; finally, leukocytes and tumor cells are distinguished based on cell morphology using a fluorescence microscope or fluorescence scanner. This invention is the first to detect the number and type of circulating tumor cells (CTCs) in peripheral blood based on ionizing radiation and radiosensitization effects, according to the degree of DNA damage in tumor cells. Compared with existing technologies, this detection method and kit are not only simple and rapid, but also significantly improve detection sensitivity and specificity. Furthermore, it is independent of the physical properties of cells and tumor cell surface markers, requires a small sample volume, is highly versatile, and can detect CTCs in different types of solid tumors.

The invention relates to the technical field of hepatocellular carcinoma detection, and discloses a postoperative circulating tumor cell detection method based on hepatocellular carcinoma. The method comprises the following steps: collecting a venous blood sample within half an hour after a hepatocellular carcinoma patient, treating the venous blood sample with a red blood cell lysis solution to remove red blood cell components, and performing centrifugal separation to obtain a cell precipitate; embedding the precipitate in a paraffin matrix to form a cell paraffin block, and continuously slicing to obtain a slice sample to be detected; labeling surface antigens of the circulating tumor cells in the slices through specific antibodies by adopting an immunohistochemical technology; collecting a microscopic image of the marked sample, and extracting morphological characteristic parameters and dyeing intensity distribution data related to the circulating tumor cells; a recognition model is constructed based on the data, and positive circulating tumor cells and background interference signals are distinguished. The method is standard in process, achieves efficient detection of circulating tumor cells, and meets clinical requirements of hepatocellular carcinoma postoperative monitoring.