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流式细胞术研究细胞学机制:"cell-cell contact"
HM16 2016-9-8 20:50
简介 细胞间的相互作用方式大体上可以分为两种,一种是通过分泌细胞因子,趋化因子等可溶性的蛋白质相互作用,这个过程不依赖于细胞间接触;另一种是依赖于细胞间接触,通过细胞表面的配体与受体结合,传导信号最终起作用。 细胞间接触机制的研究方法也比较固定:首先,体外可用Transwell实验 培养 两种细胞,阻止细胞-细胞接触,进而观察细胞-细胞接触对于该生物学现象的作用;其次,体内观察到这种细胞-细胞接触的现象;最后,干扰某个/些分子,破坏细胞-细胞接触,进而改变该生物学现象。 实验小白今天就为大家介绍一下如何巧用流式细胞术研究细胞-细胞接触机制。 例1:Nat Immunol 2009“B cell–T cell conjugates” 大量的研究表明 B细胞诱导Tfh细胞产生的过程是依赖于细胞-细胞接触机制的。2009年Nature杂志上发表的文章 就用流式细胞术证明了这一机制。 Reinhardt RL, Liang HE, Locksley RM. 2009. Cytokine-secreting follicular T cells shape the antibody repertoire. Nat Immunol 10(4):385-393. 第一步:体内发现CD19+CD4+ cells 众所周知,CD19阳性的细胞是B细胞,而B细胞表面是不表达CD4或者CD3分子的。我们现在观察到CD4+CD19+的细胞,初看好像很不合理。仔细分析这篇文章的材料方法,小鼠脾脏和淋巴结取出后,机械性研磨、裂解红细胞、流式染色过程中都未加入EDTA,而且圈门过程也未去除双联体细胞。因此,这群CD4+CD9+cells有可能就是B cell-T cell conjugates。 第二步:分析CD19+CD4+细胞大小(也就是FSC的值) 如果CD4+CD19+细胞真的是T cell-B cell conjugates,那么这群细胞的大小应该是CD4+T细胞大小和CD19+B细胞大小之和。如上图所示,原始文献中是这样描述的:Further confirming their identification as B cell–T cell conjugates, cells that expressed both T cell and B cell markers were twice the size of single cells。 第三步:分选CD19+CD4+ cells,加入EDTA打破细胞间连接,然后流式验证是CD19+B细胞和CD4+T细胞。 文献中是这样描述的:The resulting single cells segregated into approximately equal numbers of CD4+ and B220+ single-positive populations, consistent with their original purification as B cell-T cell conjugates. PS: Dual-reporter 4get-KN2 mice (IL-4 reporter). 例2:PLoS Pathog 2014“Macrophage-T cell contact” 看完上面的例子实验小白被吓到了,流式还真的能这么用啊?我们也尝试一下,照葫芦画瓢 。 Chen X, Yang X, Li Y, Zhu J, Zhou S, Xu Z, He L, Xue X, Zhang W, Dong X, Wu H, Li CJ, Hsu HT, Kong W, Liu F, Tripathi PB, Yu MS, Chang J, Zhou L, Su C. 2014. Follicular helper T cells promote liver pathology in mice during Schistosoma japonicum infection. PLoS Pathog 10(5):e1004097. 1. 体内存在F4/80+CD4+ cells 和例1类似,我们发现小鼠脾脏中也有F4/80+CD4+ cells,而且这群细胞绝大多数都表达CD3分子,初步提示这群细胞是巨噬细胞-T细胞的结合体。 2. 流式检测F4/80+CD4+细胞大小(FSC) 如上图所示,我们发现F4/80阳性的巨噬细胞大小为300,CD4+T细胞的大小为200,而F4/80+CD4+细胞的大小为500。进一步提示F4/80+CD4+ cells 很有可能是巨噬细胞与CD4+T细胞的结合体。 3. 分选F4/80+CD4+cells,用EDTA破坏细胞间粘连后,流式确证改双阳性细胞是巨噬细胞与CD4+T细胞的结合体。 流式分选出F4/80+CD4+ cells,用EDTA处理后,流式检测的结果显示基本上是等比例的F4/80+CD4-CD3-macrophage和F4/80-CD3+CD4+T cells。
个人分类: 研究方法|7737 次阅读|0 个评论
New Lab is open
热度 3 abide 2014-3-20 22:01
New Lab is open, free access, reserve seats for future postgraduates. Welcome start your study in emei campus SWJTU. Free contact me by email.(kmwang@swjtu.edu.cn) 由衷感谢几位老师的支持,新据点开始这周运营了,网络通了,水也有得喝了,可以好好着手推进项目完成了。 虚位以待,呼唤用3年时间到实验室学习钻研、改变人生的学子。 招生主页: http://userweb.swjtu.edu.cn/Userweb/abide/index.htm 陪了5年的笔记本放在身边才踏实——时间去哪了?凝神 , 聚力是第一步。 窗外是运动场,这风景不错。——就是高了点,157阶乘以6的运动量。 实验室第一位研究生张同学的工作位 用想象力改变生活,是企业的使命,也是我们前行的任务。
个人分类: 科研|24418 次阅读|3 个评论
[转载]最好的科学电影:接触(Contact)
热度 2 seawan 2013-9-25 18:07
http://player.pptv.com/v/wrJ591icFNXPWVLw.swf 两年后才知道茱蒂福斯特。 多年后又看到里面的瞎子到了《越狱》。。。 --- 《超时空接触》是目前 好莱坞 所拍摄的科学含金量最高的真正意义上的科幻片,调查表明,美国有97%的民众相信影片所述的内容。与之相比,那些全球排行榜上名列前茅的电影,如《 星球大战 》系列、《未来战士续集》及《 黑客帝国 》等,过于天马行空,只能算是狂想片。由于该片系当今好莱坞一群超级精英制作,剧情丰满,表演精湛,视听效果上乘,全球票房高达1.66亿美元。 ——百度百科 经典对白 1. Dr. Kent Clark: Nice to smell you again, Mr. Kitz. 1. 博士肯特·克拉克:很高兴再次嗅到你, Kitz先生。 2. Executive: We must confess that your proposal seems less like science and more like science fiction. Ellie Arroway: Science fiction. You're right, it's crazy. In fact, it's even worse than that, it's nuts. You wanna hear something really nutty? I heard of a couple guys who wanna build something called an airplane, you know you get people to go in, and fly around like birds, it's ridiculous, right? And what about breaking the sound barrier, or rockets to the moon? Atomic energy, or a mission to Mars? Science fiction, right? Look, all I'm asking is for you to just have the tiniest bit of vision. You know, to just sit back for one minute and look at the big picture. To take a chance on something that just might end up being the most profoundly impactful moment for humanity, for the history... of history. 2. 执行:我们必须承认,你的建议似乎不像科学,更像是 科幻 小说。 爱丽博士:科幻小说。你说得对,这太疯狂了。事实上,比这还要糟糕,这简直就是神经病式幻想! 你想听一些真正疯狂的事情吗? 我听说两个人要建造一种叫做飞机的东西,你知道,那种你让人们坐进去,像鸟一样满天飞的东西, 荒谬可笑, 对吗? 还有超音速行驶,或者乘火箭到月球上去呢? 原子能,火星任务? 科幻小说,对吗? 你看,你只需要坐回去再看一分钟这张图。你要在历史上抓住一个机会,为那些最后可能对人类,对历史有重大影响的事情…… 3. Young Ellie: Dad, do you think there's people on other planets? Ted Arroway: I don't know, Sparks. But I guess I'd say if it is just us... seems like an awful waste of space. 3. 小艾莉:爸,你认为其他行星上有人吗? 泰德?亚若威:我不知道,史巴克。但是我猜,我想说的是,如果这里仅仅只有我们…看起来就太浪费空间了。 4. Rachel Constantine: Twenty million people died defeating that son of a bitch Hitler, and HE'S OUR FIRST AMBASSADOR TO OUTER SPACE! 4. 拉 康斯坦丁 :二千万人死于打败那狗娘养的希特勒,如今他居然成了我们到外层空间的第一位大使! 5.Ellie Arroway: Mathematics is the only true universal language. 5.Arroway: 数学是宇宙中唯一真正通用的 语言 。 6.David Drumlin: Ellie, still waiting for E.T. to call? 6.Drumlin: Ellie,你还在等外星人的电话么? 7.Ellie Arroway: (listening to the message) Those are primes! 2,3,5,7, those are all prime numbers and there's no way that's a natural phenomenon! 7.Arroway:(听着信号)这些是素数!2,3,5,7,都是素数。这绝对不是正常现象! 8.Young Ellie: (on HAM Radio) CQ, CQ, this is W9GFO. Is anybody out there? 8.小Ellie:(无线电)CQ, CQ, 我是W9GFO。有人在么? 9.Ellie Arroway: Because I can’t… I... had an experience. I can't prove it, I can't even explain it, but everything that I know as a human being, everything that I am…tells me that it was real. I was given something wonderful, something that changed me forever. A vision of the universe, that tells us undeniably, how tiny, and insignificant and how... rare, and precious we all are! A vision that tells us that we belong to something that is greater than ourselves, that we are *not*, that none of us are alone. ...I wish I could share that....I wish...that everyone,if even for one...moment...could feel that awe and humility and hope...that...... That continues to be my wish. 9.Ellie:因为我不能...我有这种经验,我不能证明,也不能解释它,但是凭着我所有的知识和感觉我知道这是真的。我收获了宝贵的东西,它永远地改变了我。宇宙的景象毫无置疑地证明了我们人类是多么渺小、微不足道……但同时也是多么珍贵!这景象还告诉我们一些更伟大的事情,那就是我们不是宇宙的孤独一员。...我希望能够和你们分享我的经历。...我希望...每个人,一刻...也好...能够感受到那种敬畏,谦逊,和希望...那种......这仍然只是我的愿望。
个人分类: 博览|2346 次阅读|3 个评论
Electrical Properties of Self-aligned in-situ Reduced Graphe
zhengqingbin 2013-1-16 08:32
This mail is sent by ConfSys/ICCM system. Please don't reply. A copy of this email is in your Inbox in ConfSys. This mailbox would be accessible once you have completed your profile and topics of interests. For any ICCM paper related issues contact the event organizers. Please include your Paper ID and title. Message from: Confsys System Message to: conf909859 Dear user, This mail is to inform you that you have been registered as a co-author of a paper. The detail information for this is as follows: PID: YOU81486 Paper Title: Electrical Properties of Self-aligned in-situ Reduced Graphene Oxide/Epoxy Nanocomposites Contact(submitting) author: Nariman Yousefi Conference: ICCMICCM19 Your email address as registered in ConfSys is: qingbin.zheng@family.ust.hk Your username is: conf909859 Your password is: 408902 If there are any issues with the paper etc. please communicate with the submitting (contact) author. You would have access to the full functionality of ConfSys after you complete your profile. To do this, please access the ConfSys website: http://confsys.encs.concordia.ca/ICCM/jsp/index.jsp and login with your username and password and update your profile and interests. If you wish, you may change the password; in that case, please make a note of it. You may consult the On-line help for more details about this process PLEASE NOTE: Your registration as a user of ConfSys is not complete until you complete the registration process. ConfSys Privacy policy: ConfSys does not release the information of its users to a third party. Sincerely, Admin: CINDI/ConfSys The system displays your current event at the top of the web page, under the graphics. To access the required functions for an event, please make sure to change to the appropriate event using the Change Event link located next to the link to return to the Homepage under the graphics for ConfSys. Unless you have changed to the appropriate event, you would not be able to access the required functions for that event. Pl. use the link to get context sensitive help! The URL for ICCM site on ConfSys is: http://confsys.encs.concordia.ca/ICCM
个人分类: 科研笔记|1 次阅读|0 个评论
Ming Liu's doctoral dissertation on contact deformation
hasanzhong 2012-12-21 01:33
http://uknowledge.uky.edu/cme_etds/15
个人分类: Paper|3592 次阅读|0 个评论
骗子越来越多?
ljxm 2012-3-31 08:35
如下是我收到的邮件 是不是骗子 请鉴定 Any classes, audiences, textbooks. Bachelors, Masters and MBA and/or Doctorate (PhD) .Moreover you don抰 need to go to university and suffer all those boring long lectures! Well, that can be real! And moreover you don抰 need to study over there! Want to know more about that? WISH TO DOUBLE YOUR INCOMES? Feel interested already? Want to know more? Then contact us right now! We are ready to start to be engaged in your business immediately, Improve your qualification. Confidentiality assured! Your calls are accepted round the clock Inside USA: 1-603-509-2001. Call Outside USA.: +1-603-509-2001 Then contact us or leave a voicemail with your NAME and contact TELEPHONE NUMBER. (with country-code)
2250 次阅读|0 个评论
[转载]Contact (1997)
seawan 2011-12-3 12:32
【按:电影中文名很俗:《超时空接触》;原著: 卡尔·爱德华·萨根 (Carl Edward Sagan,1934年11月9日-1996年12月20日), 美国 天文学家 、天体化学家、科幻作家,和非常成功的天文学、天体物理学等自然科学方面的科普作家。 行星学会 的成立者, 研究 天文生物学 的先驱,也是搜寻地外智慧生物项目( SETI )的创始人之一。 。 。】 【再按:感概于美国科幻的深厚底蕴。实在不应该叫“科幻”,应该就“科学小说”或者“科学故事”,SF。】 豆瓣上的电影简介:    艾莉·爱罗维(茱迪·福斯特饰)自幼就对无线电通讯情有独钟。长大后,她成为一名天文学家,其项目内容为接收外太空信号。她希望能借此找到宇宙其他文明的存在。天才的理想得到的却是投资者的冷眼相看,幸好有神秘人帮助艾莉继续工作。   一次偶然的机会,艾莉收到了来自织女星的频率信号,似乎证明了艾莉一直以来的工作是正确的。但这带来的不是她个人事业的成功,而是政治、社会、宗教、国家各个角度的矛盾与讨论。深陷这一切暴风中心的艾莉,更要面对与神职人士帕尔默·乔丝(马修·麦康纳饰)的感情纠葛。   个人与世界,真相与现实,艾莉面临着远远超越学术范围的挑战。 http://seawan.nyist.net/kle/download.php?id=76  【经典对白】   1. Dr. Kent Clark: Nice to smell you again, Mr. Kitz.   2. Executive: We must confess that your proposal seems less like science and more like science fiction.   Ellie Arroway: Science fiction. You're right, it's crazy. In fact, it's even worse than that, it's nuts. You wanna hear something really nutty? I heard of a couple guys who wanna build something called an airplane, you know you get people to go in, and fly around like birds, it's ridiculous, right? And what about breaking the sound barrier, or rockets to the moon? Atomic energy, or a mission to Mars? Science fiction, right? Look, all I'm asking is for you to just have the tiniest bit of vision. You know, to just sit back for one minute and look at the big picture. To take a chance on something that just might end up being the most profoundly impactful moment for humanity, for the history... of history.   3. Young Ellie: Dad, do you think there's people on other planets?   Ted Arroway: I don't know, Sparks. But I guess I'd say if it is just us... seems like an awful waste of space.   4. Rachel Constantine: Twenty million people died defeating that son of a bitch Hitler, and HE'S OUR FIRST AMBASSADOR TO OUTER SPACE!
个人分类: 电影|2435 次阅读|0 个评论
一种可能的缓解学生压抑的方法:兼职Confidential contact
热度 1 gutbahn 2011-12-3 00:15
刚刚读了关于华中科技大学学生跳楼的报道, http://news.sciencenet.cn/htmlnews/2011/11/256169.shtm ,替学生可惜,作为一名老师,自己也反思了一下大学能不能有所改进。 记得以前收到过一封邮件,项目的秘书发的(德国),说一个青年博士将担任项目中学生们的confidential contact person,希望同学对与任何项目教学和科研过程中的疑问,抱怨都可以先找这个博士谈,他会全部保密。他如果解决不了,学生可以再向系里反映情况。 这个博士教过我一些编程,算是有些了解,所以我觉得这个人选特别好。首先,他是荷兰人,和德国人比起来更easy going。而且,他业务非常出色,技术上和学术思维能力都很强。还有一点,我个人感觉他挺不得志的,博士毕业十年了,还是assistant professor的岗位,独在异乡,换过两次东家,恐怕如果论苦,一般人的也敌不过他的苦闷了,所以什么大不了的心结,在他这恐怕都会化解了。 在国内,辅导员是和学生接触的一线工作人员,非常辛苦。但是,恐怕真有什么问题,学生很少会找辅导员谈心,因为辅导员上对书记负责,下要分配很多学生眼中的“利益”,谈的不好,有可能会引发“会商”之类的干戈,或者从此被打成学生中的“异类”不得重用。所以学生很多时候会找比较喜欢的老师去谈心,但是,严格上讲,这种谈心既有兄弟义气的成分,也有学生对老师的一丝忌惮,效果是否真好,也就不得而知了。所以,如果真有一个独立的不需要对任何人负责的、且能够为学生完全保密的联系人的存在,恐怕对学生应该是个福音了。而且,一个相同专业的青年博士的建议,也许和学生本身的经历更有契合之处。 希望学生们能够更快乐的学习!
3533 次阅读|2 个评论
2011年生理学和医学诺贝尔奖预测
热度 1 xupeiyang 2011-10-4 14:29
根据被引用次数预测生理学和医学诺贝尔奖获得者。 汤森路透是唯一采用定量数据预测年度诺贝尔奖得主的机构,自2002年起,共有21位引文桂冠奖得主赢得诺贝尔奖。 根据研究人员所发表论文在过去二十年间被引用的次数,汤森路透引文桂冠得主通常名列其研究领域科研人员的前千分之一。 http://news.sciencenet.cn/htmlnews/2011/9/253017.shtm 2011年生理学和医学诺贝尔奖得主Tim R. Mosmann(预测) 2011-9-23 13:06 Tim R. Mosmann, Ph.D. Contact Information University of Rochester School of Medicine and Dentistry 601 Elmwood Ave., Box 609 Rochester, NY 14642 Appointment: (585) 275-URMC Professional Bio Dr. Tim R. Mosmann is Director of the Human Immunology Center (HIC), and al ... 个人分类: 诺贝尔奖 | 234 次阅读 | 没有评论 2011年生理学和医学诺贝尔奖得主Robert L. Coffman(预测) 2011-9-23 12:57 Robert L. Coffman (vice president and chief scientific officer, Dynavax Technologies) and Timothy R. Mosmann (University of Rochester Medical Center immunologist) “for their discovery of two types of T lymphocytes, TH1 and TH2, and their role in regulating host immune response” ... 个人分类: 诺贝尔奖 | 140 次阅读 | 没有评论 2011年生理学和医学诺贝尔奖得主Miller, Jacques F A P(预测) 2011-9-23 12:49 Miller, Jacques F A P From Faculty of Medicine Online Museum and Archive Jump to: navigation , search BSc Med 1954 MB BS Hons 1956 PhD (Lon) MD Hon (US) DSc (Lon) BA (Melb) Jacques Miller, Photo courtesy of Walter and Eliza Hall Inst ... 个人分类: 诺贝尔奖 | 131 次阅读 | 没有评论 2011年生理学和医学诺贝尔奖得主Joseph P. Vacanti(预测) 热度 3 2011-9-23 12:39 Joseph P. Vacanti, M.D. John Homans Professor of Surgery, Harvard Medical School Surgeon in Chief, Massachusetts General Hospital for Children Deputy Director, Center for Regenerative Medicine, Massachusetts Gen ... 个人分类: 诺贝尔奖 | 1184 次阅读 | 2 个评论 2011年生理学和医学诺贝尔奖得主Robert S. Langer(预测) 2011-9-23 12:30 Robert S. Langer is the David H. Koch Institute Professor ( there are 14 Institute Professors at MIT ; being an Institute Professor is the highest honor that can be awarded to a faculty member). ... 个人分类: 诺贝尔奖 | 144 次阅读 | 没有评论 2011年生理学和医学诺贝尔奖得主Charles L. Sawyers(预测) 2011-9-23 12:13 Charles L. Sawyers, MD Chair, Human Oncology and Pathogenesis Program Office Phone: 646-888-2594 Office Fax: 646-888-2595 E-mail: martinb@mskcc.org Lab Phone: ... 个人分类: 诺贝尔奖 | 128 次阅读 | 没有评论 2011年生理学和医学诺贝尔奖得主Nicholas Lydon(预测) 2011-9-23 12:02 Nicholas Lydon Nicholas Lydon Institutions Amgen Alma mater University of Leeds , University of Dundee Knownfor Gleevec Notable awards Lasker Clinical Award (2009) 2009年拉斯大克临床医学奖获得 ... 个人分类: 诺贝尔奖 | 141 次阅读 | 没有评论 2011年生理学和医学诺贝尔奖得主Brian J. Druker(预测) 2011-9-23 11:30 Brian J. Druker,M.D. Specialty: Cancer and Blood Disorders Special Interest: Chronic Myeloid Leukemia Accepting Patients: Yes Languages: English Brian J. Druker Professor of Medicine, JELD-WEN Chair of Leukemia Research, and Director, OHSU Knight Cancer Institute, Oregon ... 个人分类: 诺贝尔奖 | 186 次阅读 | 没有评论
个人分类: 诺贝尔奖|5447 次阅读|1 个评论
[转载]十种与老板更好共事的方法--转载Calvin Sun, Attorney at Law
quyonggang 2011-9-17 05:21
大家好, 你肯定了解,关于工作,老板的关系很要紧的。如果老板的关系不好,你也许可以是突然失业的。由于失业的情况,你很快也是可能的有移民身份的问题。 所以,我希望,我的文章,关于怎么跟老板好的合作,帮助你。不好意思,本来是英文的。但是我感谢我的朋友,江越庭,因为他 为我翻译。 他从天普法学院毕业了,收到法律硕士,现在在University of Kansas. 下面的是英文,和中文的文章。 谢谢。 If you have questions, please do not reply to this message. Instead, contact me at csun@calvinsun.com Calvin Sun, Attorney at Law 孫自成,律师 1776 E. Lancaster Ave. Paoli, PA 19301 610-296-3947 Pr: 17 15, Ex. 22:21 箴言17:15 记住你的老板可能有敏锐的洞察力 Version 1.0 十种与老板更好共事的方法 December 18, 2007 By Calvin Sun 我的薪水用小时算 , 但是我每一分钟都在老去 老板把我快到逼到极限 我也想对他大声嚷嚷 好吧,我想今天就到此为止 … — 阿兰 · 杰克逊 , 吉米 · 布菲特 , 此时下午五点整 老板 : 你没有办法和他们一起生活,但是离开他们你又没有办法生活 . 不管喜欢不喜欢 , 我们大部分人都要和老板打交道 , 而我们打交道的方式不仅影响了我们的职业生涯和工资,更会影响我们的心理健康。下面是一些如何更好地和老板相处的建议。 觉得你有一个一窍不通的老板 ? 记住马克吐温的话 , 他曾经说过,他 14 岁的时候 , 他的父亲是令人无法容忍的愚蠢 . 接着,当他 21 岁的时候,他惊奇地发现他父亲在 7 年里学到了如此多的东西。你的老板也许比你想的要聪明 , 也许在你职业生涯的后期 , 你也会认识到这个事实 . 不管怎样,一个糟糕的老板也能给出一些好的建议 . 我记得几年前一个老板和我谈起工作的时候,他说我应该更有闯劲,主动找出人们需要的是什么而不是坐在那里等着别人下任务。 这样子想 : 你从糟糕的老板那里也可以学到一些东西 . 分析什么样的老板是糟糕的老板,然后如果当你自己成为老板的时候就可以避免这样的行为。就好像犬儒主义者提醒我们的那样 , 就算是坏掉的钟一天也会正确两次 . 明确老板的目标 软件设计师常常要考虑 “ 可追溯性 ” 。 对于软件系统来说,这一要求必须直接或者间接地和公司的目标联系在一起。理论上说,任何缺乏可追溯性的要求都是无关的,必须被排除。 同样 , 广义上来说,你要知道老板期望你做的事情(看我的下一个建议)。 但同时,你要知道你的工作如何可以帮助老板。确保你所做的,不仅仅是你自己份内的事,而且可以帮助老板达成他的目标。 知道你的老板期望从你身上获得的东西 当我年轻的时候 , 我有一次向我的母亲抱怨说我无事可做。 “ Calvin” ,她回答道, “ 你为什么不练练钢琴呢? ” 那是我最后一次向她抱怨这个事情。 如果你是个小孩的话,忽略你父母的愿望没有什么大问题。但是忽略 ( 有意或无意 ) 你老板的要求就会毁了你的事业 . 你如何能指望一个很好的绩效评估如果你根本不知道你要被如何考评?如果你知道了目标,他们是否可以被量化?如果可以,你和你的老板在评估的时候会轻松很多 . 偶尔要和老板交流一下你在做的和已经完成的事 , 确认你的老板有相同的想法 . 如果老板对你的工作有看法,那么早知道比晚知道好。至少你还有时间来做出调整。 理想状态下 , 在绩效评估的时候不应该有意外情况发生 . 如果是这样,要不然老板没有很好地让你知道工作目标,要不然你没有理解好。不要让它发生在你身上。 十种与老板更好共事的方法 不需要关注 不要成为 “ 问题雇员 ” ,那种总是需要老板检查和关注的人。相反,试着成为老板可以依靠的人。这也许不会一下子就立竿见影,但是一个好的老板会发现并欣赏这一特点。 你在工作中会做到完美吗?当然不会 . 你至少会犯一个错误或者有一问题。然而,当问题发生,你去找老板(你也必须去,就像下面说的那样)不要仅仅带着问题去 . 想一些解决问题的办法,并且准备好向老板提供你的建议。 不要让老板吃惊 不要让老板对坏消息一无所知 . 换 句话说,告诉老板如果你出现个问题或者是犯了个错误。坏消息最好是从你的嘴里说出,而不是客户、同事,更决不应该是老板的老板。你有没有和素质很差的打电 话者或者客户争执过?通话一结束,马上打电话简要报告你的老板。告诉老板你和谁说了话,为什么那个人生气,老板有可能会从那个人那里听到什么。同时要告诉 老板你的想法。 建议同样适用于好消息。让老板知道你的成功。 要不然,你的老板可能在他的老板向他祝贺的时候对此还一无所知。 成功的时候感谢老板 重要时刻终于来临 : 你站在你们团队前面 , 从你的老板或者你老板的老板那里接过奖励或者是别的什么。这时你应该要做的事情是感谢所有帮助你成功的人,尤其是你的老板。如果你的老板真的帮到你了,这很容易。但如果是那些 “ 不好对付 ” 的老板呢?你也要说些什么,当然你最好也必须保持诚实。 还记得我们之前讨论过的吗?即使是糟糕的老板也可能提供好的见解。你的老板是不是让你沮丧,使事情更难办 ? 也许,在这种情况下,你可以感谢老板帮助你 看清问题 或者是 “ 保持理智 ” 或者是 “ 帮助你从多个角度看问题 ” . 不要给人家难堪,要不然你会听起来很狡猾或者是不诚实。不管怎样,说一些关于老板帮助你的话。 不要把批评看作人身攻击 因为我们大多数人都工作都很投入 , 以至于很难把我们和工作分开 . 所以当有人批评我们的工作的时候,我们会觉得那是人身攻击。这样的反应会阻碍我们的工作进度。下一次你的老板(或者其他的任何人)批评你的工作,试着假装这个工作是别人做的。然后,从第三者的角度看这个工作,再想想批评是否有道理。 你的老板也有老板 我 们刚才讨论了明确老板的目标的重要性。同样,要知道你的老板也有老板。基于这个事实,你可以和你的老板建立起一种合作关系,因为你们两个人都有一个共同的 目标就是让你老板的老板高兴,并使得你的老板得到赞赏。建议这样的合作关系会给你的老板留下更好的印象,甚至可以让你老板的老板留意你。 一 个聪明的老板会意识到你的成功是关系到他自己的成功。所以,你做的好坏和老板利息相关。进一步来说,老板的批评也是他对你有很高期望的表现。当我刚开始工 作的时候,我很不高兴因为老板给了我一个在我看来是过难的任务。我和我父亲的一个也在同一领域工作的朋友讨论这件事情。虽然已经好几年前的事情了,我仍然 记得他的建议。 Calvin, 他说 , 给了你这个任务因为他觉得你可以把工作做好。 十种与老板更好共事的方法 不要抢老板的风头 抢老板的风头会让你的职业受到很大限制 . 所以,在公众场合纠正老板错误一定要小心。有人有曾经这样对我父亲过。当他对一个团队演讲的时候,他提到了伍斯特理工学院 . 演讲中,他正确的把它的发音说为 伍斯特 . 这个人突然间大声说 , Wellington, 你错了 . 那个是 ‘ 伍 - 切 - 斯特 .' 幸运的是,我父亲很聪明 , 用下面的回答把话岔开 : 不好意思,请原谅。英语只是我的第五语言 . 我的父亲幽默的化解了这个情形 . 然而,时隔这么多年后我还听到这个故事,这就说明了我父亲是怎么看待那个人和他所做的事。 有一种情况是可以在公众场合下纠正老板错误的 : 当你的老板误以为自己犯了错误,但实际上没有 . 假设你的老板在演讲的时候引用了一个数据。他然后停下来,并说: “ 不好意思,我想我错了。 ” 如果你知道老板其实没有错,那么完全可以打断他说: “ 不,(老板的名字),你是正确的。 ” 必要的时候管理你的老板 想 在职业生涯中进步不能够仅仅坐在那里等待任务。你必须主动地寻找机会,去解决问题。为了要达成这个目标,你要利用你老板的组织能力。向你的解释你的计划为 什么是一个良好的商业决定。然后,请求老板帮你解决官僚系统上的繁文缛节,必要的时候为你介入解决。这样,你尊重老板的地位,但是你在引导老板,使他的资 源可以为你所用。 Calvin Sun 在有关客户服务、交流和领导的组织中工作。除了这个专栏外,他还为 TechRepublic 的帮助博客写文章。 他的电子邮件地址是 csun@calvinsun.com . http://www.techrepublic.com/blog/10things/10-ways-to-work-better-with-your-boss/284?tag=content;siu-container 10 ways to work better with your boss By Calvin Sun December 18, 2007, 11:00 AM PST Takeaway: I’m gettin’ paid by the hour, and older by the minute My boss just pushed me over the limit I’d like to call him somethin’ I think I’ll just call it a day… – Alan Jackson, Jimmy Buffett, “It’s Five O’clock Somewhere” Bosses: You can’t live with them, and you can’t live without them. Like it or not, most of I’m gettin’ paid by the hour, and older by the minute My boss just pushed me over the limit I’d like to call him somethin’ I think I’ll just call it a day… – Alan Jackson, Jimmy Buffett, “It’s Five O’clock Somewhere” Bosses: You can’t live with them, and you can’t live without them. Like it or not, most of us must deal with a boss, and the way we do so affects not just our career advancement and our salary, but also our mental well-being. Here are some tips on how to get along better with your boss. Note: This information is also available as a PDF download . #1: Remember that your boss just might have useful insights Think you have a clueless boss? Remember the words of Mark Twain, who once said that when he was 14, his father was so stupid it was unbearable. Then, he continued, when he became 21, he was amazed at how much his father had learned in just seven years. Your boss might be smarter than you think, and maybe later in your career, you will appreciate that fact. Regardless, a bad boss can still offer good advice. I remember what a boss from years ago told me about the workplace. He said I should be aggressive and find out what people needed done rather than sit back and wait for assignments. Think of it this way: You still can learn from a bad boss. Analyze why that boss is a bad boss and then resolve to avoid those things if you ever become a boss yourself. As the cynic reminds us, even a stopped clock is correct twice a day. #2: Know your boss’ objectives Software developers often concern themselves with “traceability.” The requirements for a software system must directly or indirectly be tied, or traced, to the objectives of the company. In theory, therefore, any requirement that lacks such traceability should be considered irrelevant and removed. In the same way, try to see the bigger picture. You need to know what the boss expects of you (see the next tip). But at the same time, you need to understand how your job helps the boss. Make sure that what you’re doing not only meets your own job description but helps the boss achieve his or her own objectives. #3: Know what your boss expects of you When I was young, I once complained to my mother that I had nothing to do. “Calvin,” she answered, “Why don’t you practice piano?” That was the last time I ever complained to her about that topic. Ignorance of your parents’ wishes may be fine when you’re a child, but ignorance (willful or otherwise) of your boss’s expectations can kill your career. How can you expect a good performance evaluation if you’re unaware of how you’re going to be measured? If you know your objectives, are they quantifiable? If so, both of you will have an easier time during your evaluation. Every once in a while, check with your boss about what you’re doing and what you’ve accomplished and make sure your boss has that same understanding. If your boss has issues with your performance, it’s better for both of you that you know sooner rather than later, so you have time to make adjustments. In a perfect world, no surprises should arise during your performance review. If they do, either your boss didn’t communicate the objectives or you failed to understand them. Don’t let that happen to you. #4: Be low maintenance Don’t be the “problem employee,” the one the boss always has to check up and follow up on. Instead, try to be the one the boss can depend on. It might not be apparent immediately, but a good boss will recognize and appreciate that trait. Are you going to be perfect in your work? Of course not. You’re probably going to make a mistake or create a problem at least once. However, when that happens, and you go to your boss (as you should, as mentioned below), try to go not just with the report of the problem. Think of some solutions and be prepared to offer your recommendations to your boss. #5: Don’t surprise your boss Don’t let your boss be blindsided by bad news. In other words, “fee up” if you created a problem or made a mistake. It’s better that bad news about you should come from you — not from a customer, not from a co-worker, and absolutely not from your boss’s boss. Did you have a negative interaction with an abusive caller or customer? As soon as the call is finished, call your boss and give a briefing. Tell the boss who you spoke with, why that person is upset, and what the boss can expect to hear from that person. Also give your side of the story. The same advice applies to good news as well. Let your boss know about your successes. Otherwise, your boss might give the impression of being unaware of them when his or her own boss offers congratulations. #6: Acknowledge your boss in your successes The moment has arrived: You’re in front of your group, receiving an award or other recognition from your boss or your boss’ boss. An appropriate thing to do at this point is to recognize the people who made it possible, in particular your boss. It’s easy to do if your boss really did help you. What about the “difficult” boss, though? You should try to say something, but at the same time you probably should be truthful as well. Remember what we discussed above — that even a bad boss can provide good insights and examples. Did your boss discourage you or make things difficult? Maybe, in that case, you could thank your boss for helping you “keep things in perspective” or for “serving as a sanity check” or for helping you “see the problem from multiple points of view.” Don’t push things, or you may start sounding cute and insincere. However, do try to say something about your boss’ help. #7: Don’t take criticism personally Because most of us are so involved with our work, it’s hard to separate ourselves from it. So when someone criticizes our work, we view that criticism as a personal attack. Reacting that way can hinder our development and our progress. The next time your boss (or anyone else) criticizes your work, try pretending that the work was done by someone else. Then, examine it as a third party would and test the validity of the criticism. A smart boss realizes that your success is tied to his or her own success. Therefore, the boss has an interest in your doing well. Furthermore, criticism from the boss could be a sign that the boss has high expectations from you. When I first began working, I was upset because my boss had given me a task that I thought was too hard. I discussed my concern with a friend of my father, who worked in the same area as I did. Though it happened years ago, I still remember that friend’s advice. “Calvin,” he said, “ gave you that task because he thinks you can do a good job.” #8: Remember your boss has a boss We discussed earlier the importance of knowing your boss’ objectives. In the same vein, be aware that your boss has a boss as well. You can use that fact to build a collaborative relationship with your own boss, because both of you have a common objective of making the boss’ boss happy and making your boss look good. Having that collaborative relationship gives your boss a better impression of you and gives you visibility to your boss’ boss. #9: Don’t upstage your boss Upstaging your boss can limit your career mobility. Therefore, be careful of correcting your boss in public, as someone did to my father once. While he was making a group presentation, he referred to Worcester Polytechnic Institute. In doing so, he correctly pronounced it as “Woo-ster.” This person spoke up, saying, “Wellington, you’re wrong. It’s ‘Woo-ches-ter.’” Fortunately, my father was smart, deflecting the comment with the following answer: “I’m sorry. Please forgive me. English is only my fifth language.” My father humorously defused the situation. However, the fact that after all these years I still hear this story tells you what my father thought of that correction and the person who made it. There’s one instance when it’s okay to correct your boss in public: when your boss mistakenly thinks he or she made a mistake but really didn’t. Suppose your boss quotes a figure while giving a presentation. He or she then stops and says, “I’m sorry, I think I made a mistake.” If you know the boss was originally correct, it’s fine at that point to interrupt and say, “No, , you’re correct.” #10: Manage your boss when necessary Getting ahead in your career requires more than just sitting back and waiting for assignments. You must take initiative, looking for opportunities and problems to be solved. In doing so, take advantage of any organizational power your boss might have. Explain to your boss your plans and why they represent a good business decision. Then, ask your boss to fight any bureaucratic battles that may arise and to run interference for you. In doing so, you recognize the boss is the boss. However, you are directing your boss, in taking advantage of pull that you possibly lack. Ge
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Note for P311 Smoller_Second order contact
zjzhang 2009-11-20 23:32
A Note for P311 Smoller_Second order contact
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