细胞技术

Introduction Prior to using lymphoid tissue cell suspensions for flow cytometric analysis and/or for in vitro functional assays it is recommended to remove red blood cells. The eBioscience 1X RBC Lysi ...

流式细胞仪常用的几种检测方法一、测定用乙醇固定的DNA的含量1、培养细胞的DNA含量的测定制备单细胞悬液于200μl的PBS缓冲液中；加入2ml预冷的70%乙醇，4℃保存；附:细胞固定的一般步骤1) 取单细胞悬液1～2×106个细胞于PBS（PH=7.2）缓冲液中；2) 300g离心5分钟，弃上清，反复两次；3) 重悬细胞于0.5ml PBS缓冲液中；4 ...

1、 取对数生长期细胞一个T75或T25瓶。2、 将培养基换成10ml DMEM(H)+10%FBS+0.1ug/ml秋水仙素的培养基，处理1~2小时。3、 将细胞培养液倒掉，马上加入3~4ml 0.1%胰蛋白酶，并立即摇晃0.5~1min。当在显微镜下看到分裂相细胞脱壁后，马上加入终止液终止消化，并将分裂相细胞收集在一个15ml的离心管中，并在1200r/min离心4min。4、 ...

一、原理培养的细胞在一般条件下要求有一定的密度才能生长良好，所以要进行细胞计数。计数结果以每毫升细胞数表示。细胞计数的原理和方法与血细胞计数相同。计算细胞数目可用血球计数盘或是Coulter counter 粒子计数器自动计数。 血球计数盘一般有二个chambers，每个chamber 中细刻9 个1 mm正方形再细刻16 个小格，深度均为0.1 mm。当chamber 上方盖上盖玻片后，每个大正 ...

1. 注意事项：1.1. 欲冷冻保存之细胞应在生长良好(log phase) 且存活率高之状态，约为80 � 90 ％致密度。1.2. 冷冻前检测细胞是否仍保有其特有性质，例如hybridoma 应在冷冻保存前一至二日测试是否有抗体之产生。不宜将冻存细胞放置在0℃～-60℃这一温度范围内过久，低温损伤主要发生在这一温度区内，是“危险温区”。1.3. 注意冷冻保护剂之品质。DMSO 应为试剂级等级， ...

1. 收到细胞株包裹时， 请检查细胞株冷冻管是否有解冻情形， 若有请立即通知。细胞株请尽速开始培养， 或立即冷冻保存(置于�70 °C， 隔夜后， 移到liq N2)。细胞复苏的原则－快速融化：必须将冻存在-196℃液氮中的细胞快速融化至37℃，使细胞外冻存时的冰晶迅速融化，避免冰晶缓慢融化时进入细胞形成再结晶，对细胞造成损害。具体操作一. 实验前准备：1.将水浴锅预热至37℃2.用75％酒精擦拭 ...

本文来自Cyagen赛业：http://www.cyagen.com.cn干细胞在培养过程中如果环境不适合其生长就会出现部分细胞的老化现象。这是干细胞培养的一个难点也是经常出现的问题。干细胞老化表现：细胞胞浆内特别是在核区附近出现黑色颗粒，表示细胞开始出现老化； 细胞胞浆内出现空泡，则表明该细胞已老化或者已经开始分化（在全部细胞中出现少数老化细胞是正常的）； 细胞立体感逐渐消失，细胞间的间隔不清， ...

1． 材料和试剂1.1培养液A ：DMEM培养液添加10%的胎牛血清（FBS）。1.2培养液B：DMEM培养液添加10%的胎牛血清（FBS）和终浓度为0.7--1μg/ml的放线菌素D。1.3 L929细胞株：鼠结缔组织纤维母细胞，来源于22天雄性C3H鼠皮下组织。1.4 结晶紫染色溶液：0。05%结晶紫 (50mg结晶紫加入20ml乙醇加蒸馏水定容至100ml)，蒸馏水稀释。1.5 脱色液 ...

本文来自Cyagen赛业：http://www.cyagen.com.cn碱性磷酸酶（ALP）是成熟成骨细胞的标志性酶，同时成骨细胞形成的钙结节也是成骨细胞的标记物。以ALP为目标物的检测方法：1Gomori钙钴法【染色原理】ALP在pH值9.4的环境下，以镁离子作为激活剂，能够把β-甘油磷酸钠水解出磷酸，磷酸与高浓度的钙盐结合形成无色的磷酸钙，再与硝酸钴作用形成磷酸钴，经硫化胺处 ...

一、形态学观察方法（一）HE染色、光镜观察：凋亡细胞呈圆形，胞核深染，胞质浓缩，染色质成团块状，细胞表面有“出芽”现象。（二）丫啶橙（AO)染色，荧光显微镜观察：活细胞核呈黄绿色荧光，胞质呈红色荧光。凋亡细胞核染色质呈黄绿色浓聚在核膜内侧，可见细胞膜呈泡状膨出及凋亡小体。（三）台盼蓝染色：如果细胞膜不完整、破裂，台盼蓝染料进入细胞，细胞变蓝，即为坏死。如果细胞膜完整，细胞不为台盼蓝染色，则为正常细 ...

Remove Human ES cells from liquid nitrogen storage tank. Fill out a freeze/thaw form.Thaw cryovial by gently swirling in waterbath until only a small ice pellet remains being careful not to completely ...

Serum Free Media for human ES cells on MEFs:can last for 7-10 daysFinal ConcentrationAmount for 250ml Stock solution80% DMEM-F12200ml20% KO Serum Replacer50ml1% Non-essential Amino Acids2.5ml1mM L-gl ...

Let human ES cells grow until the colonies are large and the cells are pretty piled up - about the time when you would normally split or even a day past that.Treat cells with 0.2 - 0.5 mg/ml dispase. ...

hES media has a two week shelf life.hES cells should be cultured in 4 6 24 48 96 well plates. Growing cells in flasks is not recommended because it is very difficult to scrape cells in flasks.hES cell ...

Method: Essentially that described by Heins et al. (1966) based upon the release of acid soluble oligonucleotides following nuclease digestion of DNA. One unit corresponds to a change in optical densi ...

DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays such as chicken chorioallantoic membrane assay it is c ...

Reagents Heparin - 1000 U/ml Ficoll-Hypaque PBS RPMI-1640 supplemented with 10 mM glutamine and 15% FBS AET (0.14M) Dissolve 1.967 g AET in 35 ml di-H2O. Adjust to pH 8.0 with 1.0N NaOH. Bring volume ...

The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green alive cells; P.I. (propidium iodide)-red dead cells 35 mm plates: to 2 ml medium or PBS add 2 ul ...

CAM ASSAYShell-less embryo cultureFertilized white leghorn chicken eggs (SPAFAS Inc. Norwich CT) were received at day 0 andincubated for 3 days at 37°C with constant humidity. On day 3 eggs were rinse ...

PREPARING CELLSBring up HUVEC and fibroblasts in M199/10% FBS/Pen-Strep (1:100) 1-2 days before beading. Switch medium to EGM-2 (Clonetics) the day before beading for HUVEC and the day before embeddin ...