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When reached logarithm growth phase, it was poured or the medium was removed. 5 mL medium containing 10 mg/L mitocin-C was added , kept at 37 ℃, and then cultured in saturated humidity warm box with the CO2 of 0.05 fraction volume for 2 or 3 hours. It was coated with 0.1% gelatin in 6-hole plate, laid for over 30 minutes, and then threw away or the medium with mitocin-C was removed, washed with phosptat buffer for 5 times so as to remove the mitocin-C. 2 mL 0.05% trypsin digestive cells were added, and it was observed under microscope. When crevice appeared, cells became round (about 2－4 minutes), digestion was stopped by adding medium of the same volume, and blew up with straws repetitively to make it into monoplast suspension. Special-used cover glass was put in the center of cell counting chamber. Cells were sucked in by glass siphon, and cell suspension flew out at bucket of up or down-sides of counting chamber, until the cover glass was filled with fluid. Living cell were inoculated at concentrations of 3×108, 5×108, 1×109 L－1 after their number counted.
选取2～5代的小鼠胚胎成纤维细胞，待其至对数生长期倒掉或吸掉培养基，加入含10 mg/L丝裂霉素C的细胞全培养液5 mL，置37 ℃，体积分数0.05的CO2饱和湿度温箱中培养二三小时，在6孔板中加入0.1%明胶包被，放置30 min以上，倒掉或吸掉含丝裂霉素C的培养基，磷酸盐缓冲液清洗5遍，尽量除去丝裂霉素C，加入2 mL 0.05%的胰蛋白酶消化细胞，镜下观察，当细胞间出现裂隙，细胞变圆时（约2～4 min），立即加入等量的全培养液终止消化，并用吸管反复吹打，使之成为单细胞悬液，在细胞计数板中央放置专用的盖玻片，用玻璃虹吸管吸取细胞，让虹吸管在盖玻片上或下侧的计数板凹槽处流出悬液，至盖玻片下被液体充满为止。
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notchboard：凹板 凹槽侧板 梯级搁板 梯形槽口板
notchblock 凹槽滑车 | notchboard 凹板 凹槽侧板 梯级搁板 梯形槽口板 | notchbreakingtest 缺口断裂试验
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notch block：凹槽滑车
notch bending test ==> 切口挠曲试验 | notch block ==> 凹槽滑车 | notch board ==> 凹板,凹槽侧板,梯级搁板,梯形槽口板