Method for simultaneously labelling collagen type IV, macrophage and neovascularisation of tumors
A technology for macrophages and new blood vessels, applied in the field of tissue immunofluorescence, can solve problems such as the inability to meet multiple staining, and achieve the effects of convenient tissue morphological analysis, simple operation steps and clear background.
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[0054] A method for simultaneously labeling tumor stroma type IV collagen, macrophages and angiogenesis, the steps of which are:
[0055] 1. Formalin-fixed, paraffin-embedded, 4 μm thick sections of gastric cancer tissue were fixed on poly-lysine-treated detachment-resistant glass slides.
[0056] 2. Preparation of tissue slices, put the tissue slices into xylene for dewaxing three times (in different vessels), 5 minutes each time, after dewaxing, put the tissue slices into absolute ethanol for 5 minutes, 95% alcohol for 2 minutes , 95% alcohol for 2 minutes and 80% alcohol for 2 minutes, rinse with running water for 3 to 5 minutes.
[0057] 3. For antigen retrieval, prepare trisodium citrate buffer with 29.41 g of trisodium citrate (Sinopharm Chemical Reagent Co., Ltd.), 1000 ml of double distilled water, 10.5 g of citric acid (Sinopharm Chemical Reagent Co., Ltd.), 500 ml of double distilled water ml to prepare citric acid buffer solution, respectively take 20.25ml of triso...
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