估计养细胞者,很多人遇到过所谓的“黑胶虫”。最近养细胞,不幸的是,也中奖了,看到培养瓶里仿佛一夜之间冒出来的密密麻麻的小黑东西,真是有苦难言,因为曾在园子里看过有关“黑胶虫”的帖子,不好对付,没有有效的杀灭方法,据说有专杀培养基,但又有什么好的方法可以消除实验室和培养箱内的污染呢?紫外照射、甲醛高锰酸钾熏蒸------这些方法不知其对“黑胶虫”的确切效果,有些对人有毒害,有没有一种方法,即能杀死“ ...
www.33ge.com `7j5cNt8l-o肿瘤细胞培养P.}分析化学论坛化学分析仪器分析分析测试色谱电泳光谱#t$d%m"?4d肿瘤细胞在组织培养中占有核心的位置,首先癌细胞是比较容易培养的细胞。当前建立的细胞系中癌细胞系是最多的。另外肿瘤对人类是威胁最大的疾病。肿瘤细胞培养是研究癌变机理、抗癌药检测、癌分子生物学极其重要的手段。肿瘤细胞培养对阐明和解决癌症将起着不可估量的作用 ...
1:接种细胞:用含10%胎小牛血清得培养液配成单个细胞悬液,以每孔1000-10000个细胞接种到96孔板,每孔体积200ul.2:培养细胞:同一般培养条件,培养3-5天(可根据试验目的和要求决定培养时间)。3:呈色:培养3-5天后,每孔加MTT溶液(5mg/ml用PBS 配)20ul.继续孵育4小时,终止培养,小心吸弃孔内培养上清液,对于悬浮细胞需要离心后再吸弃孔内培养上清液。每孔加150ul ...
Hippocampal Neuron Cultures (and mixed cortical cultures)Submitted by: Jean Siao Ph.D.Begin by timing the pregnant mouse at E17-E19 days of gestation. Have ready the following:1.Cold Hank’s balanced ...
MTT实验是检测细胞活力的实验方法,由于细胞活力与细胞数呈正相关,因此也常常用来检测细胞的增殖情况。MTT的原理:活细胞有琥珀酸脱氢酶,将MTT还原成棕褐色沉淀。由于一般介绍园子里已经很多,笔者将自己的心得按照实验流程与大家交流交流。1、培养好细胞点板。养细胞没啥好说的,如果不知道细胞如何养,那就看看相关的文献方法。如果知道了细胞的名字,就可以上www.atcc.org检索细胞的培养信息,这个网站 ...
原代培养细胞,关键是消化,我一般酶液是新配的,要足量啊(又一次还剩三排的时候,一同学把酶液碰到水浴锅,害的我借得酶液,耽误了时间,而且组间数据紊乱,最后浪费细胞和染料,大家要注意保护好酶液啊),这样每次心理都有底,而且放入水浴37度30min,甚至扔到培养箱(不建议采用)升温。消化前,用温PBS轻洗细胞,一次,然后开始消化,24孔板加400ul酶液(0.25%胰酶和0.02%edta,如果你的ed ...
一、流式细胞术发展简史 流式细胞术(Flow Cytometry FCM)是一种可以对细胞或亚细胞结构进行快速测量的新型分析技术和分选技术。其特点是:①测量速度快,最快可在1秒种内计测数万个细胞;②可进行多参数测量,可以对同一个细胞做有关物理、化学特性的多参数测量,并具有明显的统计学意义;③是一门综合性的高科技方法,它综合了激光技术、计算机技术、流体力学、细胞化学、图像技术等从多领域的知识和成果 ...
细胞凋亡的定性检测依赖于形态学观察及DNA电泳其定量检测则需借助于流式细胞检查。使用碘化丙啶(PI)染色检测DNA含量是最早出现的凋亡定量检测方法[1]。进入90年代检测DNA断裂点的TUNEL(Terminal deoxylnucleotidyl transferase mediated-dUTP nick end labeling)技术成为凋亡定量检测的主流。1995年Vermes 首次使用荧 ...
Isoelectric Focussing of Membrane Protein by Slab Gel Method (Hancock Lab) Isolation of Outer Membrane Protein from P.Aeruginosa with Octyl-Poe (Hancock Lab) Isolation of Outer Membrane Protein w ...
Chromosomal DNA Prep : cultured cells/tissue samples (Mike A Dyer)This protocol was developed for cultured cells but should be appropriate for dissociated tissues as well. CHO Chromosome Preparation ...
Chromosome Staining and Banding Technique (Primate Cytogenetics Network)Protocols for different staining method each is in great detail. Karyotype Analysis (William H. Heidcamp) Metaphase Chromos ...
・ Comparative Genomic Hybridization (CGH) CGH is a molecular Cytogenetic method of screening a tumor for genetic changes. The alterations are classified as DNA gains and losses and reveal a ch ...
・ Hiro Hirai's Primed in Situ Synthesis (Schistosoma Genome Network)The PRINS (Primed in situ) technique uses a specific primer dNTPs with Dig-11-dUTP and DNA polymerase to perform a primer ex ...
1) 传代培养细胞染色体显示法1.培养细胞:取处于指数生长期、用较大瓶皿培养的、80%~90%汇合单层培养细胞。2.加秋水仙素:使用最终浓度为0.02~0.8微克/毫升营养液,温箱继续培养6~10小时;或用低温封闭法:把培养细胞置于4℃条件下6~12小时后,再于37℃温箱继续培养6~10小时处理(加秋水仙素)。3.采集分裂细胞:可利用分裂中期细胞体变圆与底物附着不牢特点,此时手持培养瓶,左右反 ...
Annexin V 与PI双染检测细胞凋亡改进方案标本:肺泡灌洗液标本:外周血溶血后标记检测 ...
For direct gene transfer of tibialis anterior (TA) muscle in mice: It is optimal to use 6-8 week old mice (weight 19-21 gm). Females give better immune responses for the hepatitis B surface antigen a ...
Gene Targeting Outline (University of Michigan Transgenic Animal Model Core)This is a brief outline of the steps necessary to produce mice with a mutation targeted to a specific gene. These animals ar ...
Media and Solutions Required for Routine ES Cell Culture: Version 1 (PMCI) Media and Solutions Required for Routine ES Cell Culture: Version 2 (PMCI) Media for Embryo Culture and Manipulation (Bowte ...
Labeling Microtubules (Molecular Dynamics Inc.)Microtubules are involved in many aspects of cell motion including propulsion mitosis growth and organelle transport. They are composed of a- and ß-tub ...
Mitochondrial DNA Isolation from Somatic Embryogenic Cell Cultures of Larix (Kim Marshall) Construction of Organellar DNA Libraries (OGMP)Describes procedures for DNA preparation cloning into vecto ...