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Mycelium bourgeoning test of solid culturing with liquid strain showed the optimum culture condition was as follows: 4d strain age, 10% inoculation quantity, stirring inoculation mode. Mycelia would overgrow 250ml culturing bottle within 1 week. Compared with traditional technique, the result showed that solid culturing with liquid strain by this means would reduce culture period greatly.

通过液体菌种固体栽培菌丝萌发试验,确定最佳液体菌种种龄为:摇瓶菌种6d,发酵罐菌种4d,接种量为10%,搅拌式接种,以这种方式进行固体栽培,1周内即可使菌丝于250ml培养瓶种发透,与传统方法比较,大大缩短了栽培周期。

The white powder got from the culture flask wasidentified by nanobacteria monoclonal antibody immunofluorescence andelectron microscope.

培养8周后收集培养瓶底的白色粉末,用纳米细菌单克隆抗体间接免疫荧光法对其进行鉴定,并利用电镜进一步观察其形状。

The blank group, the pure chondrogenic inductor group and the group of the G ranula of P enetrating Bone and Removing Pain mixed with chondrogenic inductor. We adopted pro-culture solution, pure chondrogenic induced culture solution ( TGF-β310ug/L , Dex10-7mol/L , VitC50mg/L ) and the chondrogenic induced culture solution which included the serum of the G ranula. All groups were cultivated in 50ml cell culture bottles. The effects of the G ranula of P enetrating Bone and Removing Pain on chondrogenic phenotype differentiation of BMSCs were investigated after being cultivated for 1, 2, 3 weeks, then cells observed by inverted phase contrast microscope and immunocytochemical stain.

3将第2代SD大鼠BMSCs分为空白对照组、单纯软骨诱导剂组及透骨消痛颗粒含药血清加软骨诱导剂组,采用原培养液、软骨诱导培养液(TGF-β310ug/L,Dex10-7mol/L,VitC50mg/L)及透骨消痛颗粒含药血清的软骨诱导培养液,50ml细胞培养瓶内进行培养,1、2、3周后通过倒置相差显微镜及免疫细胞化学染色等实验技术,研究透骨消痛颗粒对BMSCs诱导成软骨细胞的影响。

Articular chondrocytes from the knee joints of the A group were separated respectively to fragments, and then digested by enzymes, both the chondrocytes and the uncomplite digested tiny chondral tissues have been cultured in vitro for about 2 weeks.

分别取A组9只兔子的双侧膝关节股骨滑车及部分髁部的软骨,在体外进行分离和消化,未完全消化的软骨组织加软骨细胞在培养瓶内一同培养2周后配制成浓度为1.6×10~8/ml的软骨细胞悬液,等分装于两个EP管中。

Amplified cells were cultured in PGC culture medium and non-adherence culture flask till idiosome formed, adhered and differentiated.

体外分化采用无黏附特性的培养瓶和除去白血病抑制因子的原始生殖细胞培养液,以脱饲养层法悬浮培养扩增的细胞,直到拟胚体形成及贴壁分化。

Methods: The fibroblasts companying human hepatoma were primarily cultured with explant culture method, first passaged after they spread to the bottom of the culture bottle, and pured by enzyme digestion and repeated strapping the cells were identificated through observing the morphologic change using invert microscope and detecting expression of vimentin, keratin by iMmunohistochemical method.

应用组织块培养法进行人肝癌伴生成纤维细胞的原代培养,细胞铺满培养瓶底后首次传代,通过胰酶消化法和反复贴壁法进行成纤维细胞的纯化;通过倒置显微镜观察细胞形态、免疫组织化学方法检测细胞膜波形蛋白、角蛋白表达情况,对细胞进行鉴定。

Results: The cultured chondrocytes were polygonal cells. There were many rough endoplasmic reticula and mitochondria in cytoplasm, and a lot of secretory vesicles under cell membrane and in the cytoplasm. When cultured for 10 days, some small and white nodules were formed on the bottom of the culture dished, and volcanic-mouth-like structures were formed when cultured for 20 days. Both these nodules and structures contained GAG-positive substances were demonstrated by safranine-O staining.

结果:体外培养的软骨细胞为多角形,透射电镜下见胞浆内有丰富的粗面内质网系统及线粒体,胞膜下及胞浆中有较多分泌泡;连续培养10天时,培养瓶底部出现肉眼可见的白色结节,蕃红-O染色显示白色结节含大量氨基葡聚糖阳性物质,20天时形成火山口样结构,也被蕃红-O染成深红色。

The second passage was obtained after the cell overgrew the bottom of the culture flask and purified by trypsinization and adherence repeatedly methods.

方法1。用组织块贴壁培养法获得原代卵巢癌CAFs和卵巢NFs,细胞铺满培养瓶后首次传代,通过胰酶消化法和反复传代法进行细胞的纯化;2。

The cells were cultured in 24-pore plate and 25cm2 plastic culture flask.

分别在24孔板和25 cm2塑料培养瓶内培养。

Methods Cells were maintained in DMEM medium with 10% fetal bovine serum. Five days before the beginning of experiments, the cells were seeded in phenol red-free DMEM medium containing 5% charcoal dextran-treated FBS. The cells were harvested, and seeded in 6-well culture plates or in 75ml flacks. After NP, BisA and DBP treatments for 72h, the cells were harvested, and mRNA and protein expression of PCNA, bcl-2 and bax were detected by reverse transcription-polymerase chain reaction and immunohistochemistry, respectively.

方法]T47D细胞在含10%胎牛血清的DMEM培养液中进行常规传代培养,实验前将细胞转移至无酚红DMEM(含5%活性碳葡聚糖苷处理过的FBS)中继续培养5d,收集细胞,PBS洗涤后接种於内置盖玻片的6孔板或75ml培养瓶中,用32×10^(-7)mol/L NP、32×10^(-7)mol/L BisA及32×10^(-6)mol/L DBP对细胞分别处理72h,用半定量RT-PCR技术观察这3种化合物对核增殖抗原PCNA、bcl-2及bax mRNA表达的影响,并用免疫组化方法对结果进行验证。

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