细胞技术

Paraformaldehyde Fixation of Cells BackgroundThis fixation method is good for cells labelled by fluorochrome-conjugated antibodies to membrane antigens. It will stabilize the light scatter and labelli ...

Fixation 1) Collect 2 X 106 cells. 2) Pellet cells by spinning at 1000 rpm 4°C for 5 minutes. 3) Resuspend cell pellet in 1 ml of cold PBS. 4) Fix cells by adding 4 ml of -20°C absolute ethano ...

From early post-implantation stages to approximately 18 weeks' gestation cytotrophoblast at the tips of anchoring villi at the placental periphery proliferate to form columns. From the distal edges of ...

Materials Chick embryo (approximately 8 days old) 70% (v/v) ethanol for swabbing Sterile scissors forceps and probes Sterile petri plates Phosphate buffered saline (PBS) Trypsin cold sterilized in a 1 ...

Whole mount staining. The protocol given uses a single primary antibody with nuclear counterstaining but it can be extended to double antibody staining. It requires an inverted microscope with fluores ...

1.Basic Techniques - The 'Do's and Don'ts' of Cell Culture Given below are a few of the essential "do's and don'ts" of cell culture. Some of these are mandatory e.g. use of personal protecti ...

How to thaw serum: Serum that is stored at -10º C to -40º C is stable for extended periods of time. It is neither necessary or desirable to store serum at -70º C as it does not prolong ...

细胞培养常见问题及解答

Introduction A major advance in our knowledge of cells came about with the ability to maintain them in continous culture. Prokaryotes have been cultured for a relatively long time but eukaryotic cultu ...

62 氯化钠结晶紫增菌液成分:蛋白胨 20g氯化钠 40g0.01%结晶紫溶液 5mL蒸馏水 1000mLpH9.0制法:除结晶紫外，其他按上述成分:配好，加热溶解。约加30%氢氧化钾溶液4。5mL，校正pH。加热煮沸，过滤。再加入结晶紫溶液，混合后分装试管。121℃高压灭菌15min。 63 氯化钠蔗糖琼脂成分:蛋白胨 10g牛肉膏 10g氯化钠 50g蔗糖 10g琼脂 18g0.2%溴麝香草酚 ...

51 三糖铁琼脂(换用方法)成分:蛋白胨 15g眎胨 5g牛肉膏 3g酵母膏 3g乳糖 10g蔗糖 10g葡萄糖 1g氯化钠 5g硫酸亚铁 0.2g硫代硫酸钠 0.3g琼脂 12g酚红 0.025g蒸馏水 1000mLpH7.4制法:将除琼脂和酚红以外的各成分溶解于蒸馏水中，校正pH。加入琼脂，加热煮沸，以溶化琼脂。加入0。2%酚红水溶液12。5mL，摇匀。分装试管，装量宜多些，以便得到较高的底层 ...

41 GN增菌液成分:胰蛋白胨 20g葡萄糖 1g甘露醇 2g柠檬酸钠 5g去氧胆酸钠 0.5g磷酸氢二钾 4g磷酸二氢钾 1.5g氯化钠 5g蒸馏水 1000mLpH7.0制法:按上述成分配好，加热使溶解，校正pH。分装每瓶225mL，115℃高压灭菌15min。 42 肠道菌增菌肉汤成分:蛋白胨1 10g葡萄糖 5g牛胆盐 20g磷酸氢二钠 8g磷酸二氢钾 2g煌绿 0.015g蒸馏水 100 ...

31营养琼脂成分:蛋白胨 10g牛肉膏 3g 氯化钠 5g琼脂 15～20g蒸馏水 1000mL制法:将除琼脂以外的各成分溶解于蒸馏水内，加入15%氢氧化钠溶液约2mL，校正pH至7.2～7.4。加入琼脂，加热煮沸，使琼脂溶化。分装烧瓶，121℃高压灭菌15min。注:此培养基可供一般细菌培养之用，可倾注平板或制成斜面。如用于菌落计数，琼脂量为1.5%;如作成平板或斜面，则应为2%。 32营养肉汤 ...

21 过氧化物酶试验试剂:2%儿茶酚溶液。3%过氧化氢溶液。试验方法:挑取固体培养基上菌落一接种环，置于洁净试管内，滴加2%儿茶酚溶液1mL及3%过氧化氢溶液1mL。静置于室温(20℃)中30～60min，观察结果。结果:阳性反应，细菌变为黑褐色;阴性反应，细菌不变色。注:过氧化物酶的作用可受氰化钾的抑制。 22 磷酸盐缓冲液储存液磷酸二氢钾 34g1mol/L氢氧化钠溶液 175mL蒸馏水 82 ...

11苯丙氨酸培养基成分:酵母浸膏 3gDI-苯丙氨酸(或L-苯丙氨酸1g) 2g磷酸氢二钠 1g氯化钠 5g琼脂 12g蒸馏水 1000mL制法:加热溶解后分装试管，121℃高压灭菌15min，使成斜面.试验方法:自琼脂斜面上挑取大量培养物，移种于苯丙氨酸琼脂，在36±1℃培养4h或18～24h.滴加10%三氯化铁溶液2～3滴，自斜面培养物上流下，苯丙氨酸脱氨酶阳性者呈深绿色。 12 ...

01 糖发酵管成分:牛肉膏 5g蛋白胨 10g氯化钠 3g磷酸氢二钠(Na2HPO4•12H2O) 2g0.2%滇麝香草酚蓝溶液 12mL蒸馏水 1000mLpH7。4制法:1葡萄糖发酵管按上述成分配好后按0.5%加入葡萄糖分装于有一个倒置小管的小试管内121℃高压灭菌15min。2其他各种糖发酵管可按上述成分配好后分装每瓶100mL121℃高压灭菌15min。另将各种糖类分别配好10% ...

Materials Growing root tips of the broad beanV. faba (2n=12) Solutions of chromosome-damaging agents listed in the introduction to this Chapter Reagents for Feulgen stain (Exercise 2.5) Oven at 60° ...

Materials Monolayer cultures grown in 75 mm culture flasks (Cells from Exercise 11.4 may be used or cultures of tetrahymena yeast or algae may be used.) H-thymidine with at least 4 µc/ml 0.36 c/ ...

Materials Fibroblast cells in log phase growth Ca Mg free-phosphate buffered saline (PBSA) 5% (w/v) Glutaraldehyde (GTA) 2% (w/v) Perchloric Acid (PCA) Subbed slides (coated with chrom alum gelatin) a ...

Materials Prepared slides of Ascaris megalocephala maturation Microscope Procedure 1.Place the slide(s) of Ascaris on the microscope and focus on the eggs within the body. Unlike the grasshopper testi ...