细胞技术

一、纺锤体阻断剂（Spindle inhibitor）在有丝分裂过程中，随着纺锤丝的形成，染色体被牵引到一起难以观察其形态。纺锤体的形成在于细胞质和纺锤体成分的粘度之间的平衡，因此，改变细胞质的粘度，即可破坏纺锤体形成，从而使得染色体均匀散开，且染色体缢痕区更为清楚。在培养中使用的纺锤体阻断剂为秋水仙素，在终止培养前加入适量秋水仙素，使正在分裂的细胞停留在中期，以获得大量分裂相供分析之用。秋水仙素 ...

PurposeThe main purpose of the flow chamber assay is to visualize and measure interactions between flowing cells expressing a given adhesion molecule on their surface and their receptor either directl ...

PurposeMaterials10ml 6% dextran + 7ml citrate/citric acid Dextran: T500 -- 6g+100ml PBS Citrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS 43 ml blood 12 ml RT Histopaque 1077 18 ml cold H ...

IntroductionLive imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows following dynamic cellular processes as they unfold "in ...

IntroductionMulticolour 3D-FISH in combination with confocal microscopy 3D image reconstruction and quantitative image analysis is an efficient tool for the analysis of the 3D genome structure and of ...

地高辛（DIG）产品选择指南――选择合适的标记方法标记DNA探针如果您有足够量的高纯度模板，建议您采用随机引物标记方法的DIG High Prime DNA Labeling and Detection Starter Kit I 和II。这两个试剂盒包含了标记、封闭、杂交和检测所需要的所有试剂，是真正意义上的一站式服务的试剂盒。二者之间的差别在于检测方法的不同：Starter Kit I采用了显 ...

Materials:Colcemid (GIBCO: cat #: 15210-040)Hypotonic solution (pre-warmed 37C): 1:1 0.4% KCl + 0.4% Sodium citrateFixative: 3:1 MeOH + Acetic acid (e.g. 15 mL MeOH + 5 mL Acetic acid)Pasteu ...

（一）实验前应明确的问题1.选择适当的细胞接种浓度。一般情况下，96孔培养板的一内贴壁细胞长满时约有105个细胞。但由于不同细胞贴壁后面积差异很大，因此，在进行MTT试验前，要进行预实验检测其贴壁率、倍增时间以及不同接种细胞数条件下的生长曲线，确定试验中每孔的接种细胞数和培养时间，以保证培养终止致细胞过满。这样，才能保证MTT结晶形成酌量与细胞数呈的线性关系。否则细胞数太多敏感性降低，太少观察 ...

Materials: RPMI 1640 medium fetal calf serum (FCS) 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents Best.-Nr. 295892) cell cuture flask Phythemaglutinin PHA-L (Seromed M 5030) CO2 cell cul ...

Materials 4 mice from each genotype 4 Ly5 mice Buckets with wet ice 3x Bucket with dry ice 1x Dewar flask with liquid nitrogen 100 mL beakers with 95% ethanol 2x 100 mL beaker with dd. H2O 2x Sterile ...

NoteThis protocol describes semi-automated cell counts using fluorescently labeled cells a hemocytometer and ImageJ software. The hemocytometer is not needed if you have already calibrated the areas ...

MaterialsWildtype MoMLV virus aliquot. Stored at -80ºC.Medium: DMEM + 10% FBSNIH 3T3 TK- cellsXC cellsPolybrene 1000x stock = 4 mg/mL sterile filtered.U.V. light source on a stand.ProcedureDay 0: T ...

Material:Packaging Cells e.g. Phoenix cells (an adenovirus Ad5-transformed human embryonic kidney cell line 293T transfected with two MoMLV packaging gene constructs: CMV-Env-PolyA and RSV-Gag/Pol-Tyt ...

（一）原理 混合单个核细胞悬液在通过尼龙毛柱时，B细胞、浆细胞、单核细胞和一些辅助细胞被选择性粘附于尼龙毛上，而多数T细胞则通过尼龙毛柱，这是获得富含T细胞群的有效方法。（二）材料及试剂1．尼龙毛（Nylon Wool Fiber）。2．烧杯、铝箔、漏斗、一次性手套等。3．装填尼龙毛柱，一次性注射器。4．取自然沉降的上层血浆，过聚蔗糖―泛影葡胺分层液后，获取的血浆和分层液之间的单个核细胞层。（三 ...

1、将转染后的细胞按照一定的比例接种到6孔板中（一般选用1：10和1：100两个梯度），分别采用1200mg/ml，1000mg/ml和800mg/ml的G418进行筛选；以转染携带EGFP质粒载体的细胞作为阳性对照，以未转染质粒载体的细胞作为阴性对照；2、每3～4天换液一次；3、筛选20～30天，即可得到稳定表达细胞株；4、传代后，以一定的G418浓度维持培养； ...

Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the complex organization of the mammalian testicular ti ...

The dissociation of adherent mesenchymal stem cell (MSC) monolayers with trypsin and enzyme-free dissociation buffer was compared. A significantly lower proportion of viable cells were obtained with e ...

1. 3流式细胞技术检测法原理：包绕着鞘液的单细胞或微粒经过流式细胞仪检测区域时被激发光激发出荧光信号，形成的不同波长荧光信号被光电倍增管接收后转换为电信号并进一步转换成计算机可识别的数字信号。其应用范围广泛，常用于细胞生物学、细胞遗传学、免疫学、肿瘤学、血液学等领域。应用：1）了解细胞大小及颗粒度2）细胞状态：pH 值、氧化还原、氧自由基、Ca2+3）细胞表面蛋白：CD 系列等4）细胞内蛋白： ...

The plasticity of differentiated adult cells could have a great therapeutic potential but at the same time it is characteristic of progression of serious pathological states such as cancer and fibrosi ...

1. 4细胞活性测定原理：根据活细胞线粒体内的脱氢酶可将试剂中的有效成分转变为有颜色的Formazan，并可被酶标仪检测，间接反应活细胞数量。对于细胞增殖、凋亡、生长受抑等可通过细胞数量间接得知的生物学改变，采用细胞活性测定可以提供可应用和参考的数据。应用：用于生物活性因子的活性检测、大规模药物筛选、细胞增殖、细胞毒性、药敏实验等一般流程：1） 细胞种于96孔板中，按照客户需要进行各种处理2） ...