流感诊断试剂盒（快检方法）

广州健仑生物科技有限公司

广州健仑长期供应各种流感检测试剂，包括进口和国产的品牌，主要包括日本富士瑞必欧、日本生研、美国BD、美国NovaBios、美国binaxNOW、凯必利、广州创仑等主流品牌。

流感诊断试剂盒（快检方法）

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【公司名称】 广州健仑生物科技有限公司
【市场部】    杨永汉

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【腾讯  】 2042552662
【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期2幢101-103室

实验步骤
1．贴壁细胞
1)   取普通洁净盖玻片于70%乙醇中浸泡5分钟或更长时间，无菌超净台内吹干或用细胞培养PBS或0.9%NaCl等溶液洗涤三遍，再用细胞培养液洗涤一遍。将盖玻片置于六孔板内，种入细胞培养过夜，使约为50%-80%满。
2)   刺激细胞发生凋亡后，吸尽培养液，加入0.5ml固定液，固定10分钟或更长时间（可4℃过夜）。
3)   去固定液，用PBS或0.9%NaCl洗两遍，每次3分钟，吸尽液体。洗涤时宜用摇床，或手动晃动数次。
4)   加入0.5ml Hoechst 33258染色液，染色5分钟。也宜用摇床，或手动晃动数次。
5)   用PBS或0.9%NaCl洗两遍，每次3分钟。
6)   滴一滴抗荧光淬灭封片液于载玻片上，盖上贴有细胞的盖玻片，尽量避免气泡。使细胞接触封片液，切勿弄反。
7)   荧光显微镜可检测到呈蓝色的细胞核。
2. 悬浮细胞
1)   离心收集细胞样品于1.5ml离心管内，加入0.5ml固定液，缓缓悬起细胞，固定10分钟或更长时间（可4℃过夜）。
2)   离心去固定液，用PBS或0.9%NaCl洗两遍，每次3分钟。洗涤期间手动晃动。
3)   zui后一次离心后吸去大部分液体保留约50ml液体，再缓缓悬起细胞，滴加至载玻片上，尽量使细胞分布均匀。
4)   稍晾干，使细胞贴在载玻片上不易随液体流动。
5)   均匀滴上0.5ml Hoechst 33258染色液，染色5分钟。用吸水纸从边缘吸去液体，微晾干。
6)   用PBS或0.9%NaCl洗两遍，每次3分钟。
7)   滴一滴抗荧光淬灭封片液于载玻片上，盖上一洁净的盖玻片，尽量避免气泡。
8)   H. 荧光显微镜可检测到呈蓝色的细胞核。

Experimental steps
1. Adherent cells
1) Take ordinary clean coverslips immersed in 70% ethanol for 5 minutes or longer, blow dry in a sterile clean bench or three times with a cell culture solution such as PBS or 0.9% NaCl and wash with the cell culture solution Again Cover glass slides were placed in six-well plates and seeded in cell culture overnight to make about 50% -80% full.
2) Stimulate the cells to apoptosis, exhaustion of culture medium, add 0.5ml fixative, fixed for 10 minutes or longer (4 ℃ overnight).
3) Remove the fixative and wash twice with PBS or 0.9% NaCl for 3 minutes each to aspirate the fluid. Washing should be shaker, or manually shaking several times.
4) Add 0.5 ml Hoechst 33258 stain and stain for 5 minutes. Also suitable shaker, or manually shaking several times.
5) Wash twice with PBS or 0.9% NaCl for 3 minutes each.
6) Drop a drop of antifluorescence to quench the sealing solution on the slide, cover the cover glass with cells, try to avoid bubbles. Make the cells in contact with the sealing solution and do not turn it off.
7) The fluorescence microscope can detect the blue nucleus.
2. Suspension cells
1) Collect the cell samples by centrifugation in 1.5 ml centrifuge tubes, add 0.5 ml of fixative solution, suspend the cells slowly and fix for 10 minutes or more (4 ℃ overnight).
2) Centrifuge the fixative and wash twice with PBS or 0.9% NaCl for 3 minutes each. Manual shaking during washing.
3) After the last centrifugation to absorb most of the liquid to retain about 50ml liquid, and then slowly suspended cells, added dropwise to the slide, try to make the cells evenly distributed.
4) Slightly dry, the cells attached to the slide is not easy to flow with the liquid.
5) Dilute 0.5 ml Hoechst 33258 stain evenly and stain for 5 minutes. Wipe off the liquid from the edges with blotting paper and allow to dry slightly.
6) Wash twice with PBS or 0.9% NaCl for 3 minutes each.
7) Drop a drop of antifluorescence to quench the sealing solution on the slide, covered with a clean cover glass, try to avoid bubbles.
8) H. Fluorescence microscopy detects blue nuclei.