83 results about "Immunofluorescence staining" patented technology

The invention discloses a method for culturing human normal lung tissue and lung cancer tissue organoids in vitro, comprising obtaining fresh human lung tissue cells and digesting them into single cells with collagenase; cultivating human lung tissue and lung cancer tissue organoids under 3D culture conditions in vitro . The method of culturing human normal lung tissue and lung cancer tissue organoids in vitro: obtain fresh human lung tissue cells, digest them into single cells with collagenase; cultivate human lung cancer tissue organoids under 3D culture conditions in vitro; H&E staining to clarify the structure and morphology , q-PCR detection of related gene expression; immunofluorescence staining to identify cell source detection of related protein expression. And the method of constructing mouse animal models based on the above-mentioned organoids is of great significance for the construction of large-scale and consistent human-derived and orthotopic lung cancer animal models, providing a good foundation for basic research on lung cancer and related clinical applications prospect.

The invention relates to an immunofluorescence staining method for suspension cells. The immunofluorescence staining method for the suspension cells is characterized by comprising the following steps of: collecting cells into a centrifuge tube; performing 800-gram centrifugation to collect the cells; respectively adding paraformaldehyde to fix, Triton-X100 to pass through and 1 percent bovine serum albumin (BSA) to close; adding a primary antibody and a secondary antibody sequentially to incubate; washing by an 800-gram centrifugation method after the operation of each step; performing diamino phenyl indole (DAPI) staining; and dripping on a glass slide and closing the glass slide to observe. The immunofluorescence staining method for the suspension cells has the advantages that: the immunofluorescence staining process is performed in the centrifuge tube, so the problems that the suspension cells grow on the glass slide difficultly and drop off easily in the test process are solved; the staining result is good; and the method is suitable for the suspension cells and cells which are adhered to the wall difficultly.

The invention belongs to the field of cell biology, and in particular relates to an in vitro three-dimensional culture model for studying vascular mimicry of glioma stem cells and its application. The preparation of the in vitro three-dimensional culture model is carried out according to the following steps: the three-dimensional collagen scaffold is soaked in DMEM medium for 6-24 hours and then taken out, and the redundant DMEM adhered to the three-dimensional collagen scaffold is removed to obtain a pretreated three-dimensional collagen scaffold. Treat the three-dimensional collagen scaffold and place it in a cell culture plate, plant the glioma stem cells into the pretreated three-dimensional collagen scaffold, let it stand at 37°C for 2-6 hours, add endothelial medium, and culture at 37°C for 2-4 days , which is an in vitro three-dimensional culture model for studying vascular mimicry of glioma stem cells. All operations of this in vitro three-dimensional culture model are carried out at room temperature, and various stainings can be performed in situ, including immunohistochemistry and immunofluorescence staining; electron microscope detection is possible; RT‑PCR and Western Blot detection can be performed directly after digestion .

The invention relates to medicine application of 20(S)-ginsenoside Rg3, in particular to application of 20(S)-ginsenoside Rg3 in preparation of medicines for treating a non-small cell lung cancer. In the invention, selecting a human non-small cell lung cancer cell strain (A549 lung adenocarcinoma, H460 large cell lung cancer and LTEP-78 lung squamous carcinoma) to be inoculated under the skin of a naked mouse, and observing the influence of the SPG-Rg3 on the non-small cell lung cancer; and carrying out CD34 immunohistochemistry staining and TUNEL immunofluorescence staining on tumor tissues,and observing the influence of the SPG-Rg3 on tumor angiogenesis and apoptosis. The inclusion proves that the 20(S)-ginsenoside Rg3 can remarkably inhibit the growth of the non-small cell lung cancer, has the action mechanism related with the promotion of the tumor apoptosis and the inhibition of the tumor angiogenesis and unobvious cooperation action when being combined with cyclophosphamide forapplication, and can be used for the clinical chemotherapy of the non-small cell lung cancer.