A nucleic acid aptamer-based immunofluorescence kit for pulmonary metastatic osteosarcoma tissue
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A nucleic acid aptamer and immunofluorescence technology, which is applied in the field of tissue immunofluorescence to achieve the effects of sensitive detection, simple operation and good tissue structure preservation.
Inactive Publication Date: 2019-05-10
FUZHOU UNIV
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[0005] At present, there are few reports on the use of nucleic acid aptamers for the diagnosis and treatment of osteosarcoma cells with lung metastasis, and the treatment of osteosarcoma faces many challenges. For example, osteosarcoma cells are very prone to lung metastasis. Therefore, there is an urgent need for an effective As a means of diagnosis and treatment, nucleic acid aptamers can provide effective research means for the diagnosis of osteosarcoma
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Embodiment 1
[0028] Detection of pulmonary metastatic osteosarcoma tissue by immunofluorescence kit based on nucleic acid aptamer
[0029] 1. Collection of tissue samples:
[0030] Sampling was performed in the operating room, and samples were taken from the osteosarcoma tissue of the same patient for the preparation of Cy5-labeled stained sections. All samples came from Mengchao Hepatobiliary Hospital of Fujian Medical University.
[0031] 2. Tissue paraffin section production:
[0032] Referring to the conventional procedure, the specific steps include fixation (overnight), elution and dehydration (6 hours), clearing (80min), wax dipping and embedding (4 hours), sectioning, patching and baking.
[0033] 3. Nucleic acid aptamer incubation for tissue paraffin sections:
[0034] (1) Bake the paraffin slices in an oven at 60°C for 2 hours to melt the wax on the surface;
[0035] (2) Dewaxing: immerse paraffin sections in xylene twice, 15 minutes each time;
[0036] (3) Hydration: Immerse...
Embodiment 2
[0046] Aptamer-based Immunofluorescence Kit for Lung Metastatic Osteosarcoma Tissue Detection of Normal Bone Tissue
[0047] 1. Collection of tissue samples:
[0048] Sampling was performed in the operating room, and samples were taken from the normal bone tissue of the same patient for the preparation of Cy5-labeled stained sections. All samples were obtained from Mengchao Hepatobiliary Hospital of Fujian Medical University.
[0049] 2. Tissue paraffin section production:
[0050] Referring to the conventional procedure, the specific steps include fixation (overnight), elution and dehydration (6 hours), clearing (80min), wax dipping and embedding (4 hours), sectioning, patching and baking.
[0051] 3. Nucleic acid aptamer incubation for tissue paraffin sections:
[0052] (1) Bake the paraffin slices in an oven at 60°C for 2 hours to melt the wax on the surface;
[0053] (2) Dewaxing: immerse paraffin sections in xylene twice, 15 minutes each time;
[0054] (3) Hydration: ...
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Abstract
The invention provides a nucleic acid aptamer-based lung metastatic osteosarcoma tissue immunofluorescence kit. The nucleic acid aptamer-based lung metastatic osteosarcoma tissue immunofluorescence kit comprises a), an antigen repairing liquid; b), a washing liquid; c), a nucleic acid aptamer dilution liquid; d), a blocking liquid. In applications, Cy5 labelled nucleic acid aptamer is taken as a fluorescence probe for labeling of paraffin embedded lung metastatic osteosarcoma tissue. The nucleic acid aptamer is capable of realizing specific combination with lung metastatic osteosarcoma cells,and combination with other cells is impossible to realize, so that the nucleic acid aptamer is taken as a novel molecule probe with high specific recognition capacity on lung metastatic osteosarcoma,and possesses important research meaning in early stage diagnosis and treatment of osteosarcoma susceptible to pulmonary metastasis. In applications, paraffin section immunofluorescence dyeing is adopted, pathological diagnosis on lung metastatic osteosarcoma tissue can be realized without low temperature storage, the specificity is high, the sensitivity is high, operation is simple, equipment requirement is low, positioning performance is excellent, and reality meaning in domestic popularization is achieved.
Description
technical field [0001] The invention relates to the technical field of tissue immunofluorescence, in particular to a nucleic acid aptamer-based immunofluorescence kit for pulmonary metastatic osteosarcoma tissue. Background technique [0002] Osteosarcoma is a kind of non-hematological malignant connective tissue tumor with a high incidence rate. It is more common in children and adolescents, especially young men. Osteosarcoma is generally caused by osteoblasts with a relatively low degree of differentiation. It has Very high lung metastasis characteristics, the tumor is very malignant, the prognosis is extremely poor, and lung metastasis can appear within a few months. [0003] Nucleic acid aptamer (aptamer), also known as nucleic acid aptamer and aptamer, refers to the method of systematic evolution of ligands by exponential enrichment (Systematic Evolution of Ligands by Exponential Enrichment, SELEX) technology from random single-stranded DNA / RNA library The screened sin...
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