n********k
发帖数: 2818 | 1 A Genetic and Functional Relationship between T Cells and Cellular
Proliferation in the Adult Hippocampus,
Sorry I remembered the opposite...I think they are working on the opposite
as the first author contacted us when my paper was out to collaborate and
dissect the opposite part... |
|
S*****3
发帖数: 720 | 2 “中国百篇最具影响优秀国际学术论文” 评选所选论文代表了我国科技论文发展的最
高水平。论文源为前一年被SCI（科学引文索引）收录的中国论文。评选综合考虑发表
论文的期刊水平（影响因子和单篇引用次数）、论文类型、热点论文、论文的合作强度
、参考文献数和论文的完整性等方面。
其中生物类包括：
论文题目: Acute promyelocytic leukemia: from highly fatal to highly
curable
论文作者: Wang, Zhen-Yi; Chen, Zhu
所属机构: 上海交通大学医学院瑞金医院
来源期刊: BLOOD, 2008, 111(5):2505-2515
被引次数: 29
作者简介：
陈竺
卫生部部长，中国科学院院士，博士生导师
白血病系统生物学研究组组长
论文题目: Sorting of small RNAs into Arabidopsis argonaute complexes
is directed by the 5 ' terminal nucleotide
论... 阅读全帖 |
|
p******i
发帖数: 1092 | 3 虽然SC说抗体能IP，建议你最好用自己的sample做个routine的IP+Western试一下。确
定能IP了，再考虑CROSS LINK？
有关重复用agarose beads的问题
1 IP需要多少抗体？没觉得省很多啊……既然用的是原代细胞，你的DISSECTION和
CULTURE的人工费用应该比抗体贵很多吧
2 要确保经过 1和DSS反应 2被ELUTION buffer摧残 后的抗体仍然好用……
浓缩洗脱组分（随便换buffer或者desalt）可以用spin column，
参见
http://www.life.sci.qut.edu.au/epping/LSB607OLT/607concentrator |
|
|
|
n***w
发帖数: 2405 | 6 你所有的染色都应该在EP管里进行。此时，retina仍然是杯状（已经从RPE分离了）。
下面你要在dissection scope下面进行，用transfer pipette (前面的尖头剪掉)将液
体吸得差不多，然后把EP管倒置，这个时候retina应该就在盖子里，然后用镊子小心地
将其夹出，放到scope下的slide上，这时，retina应该还是呈现杯状, ganglion cell
layer up.
然后你要做的就是剪4下（一般四下就够了）。随便找个地方先剪一下，这个时候张力
减小，有两个地方就应该贴到slide上了，然后在对称的地方再剪一下，这个时候，一
般整个retina都贴上去了。
然后你在这两个切口90度的地方在各剪一下，所以四个口一般足够了。
至于peripheral的地方要弄平，我习惯直接用镊子，但是鉴于每个人小心程度没有办法
衡量，你也可以用soft brush，小心的弄平。
有什么问题可以再问我。 |
|
n***w
发帖数: 2405 | 7 我的standard protocol是：
1. enucleate the eyes
2. slit the cornea, put the eye in the fixative (usually 4% PFA), for 10min
3. remove cornea and lens and vitreous, and put the eyecup (retina+RPE+
sclera, etc) back to the fixative for about 30-40min. 1hour should also work
.
4. wash with PBS and dissect the retina from the eyecup.
5. put the retina to PBSTriton (1%) permeabilize
6. staining part
这个protocol适用于~95%的蛋白染色。
但是如果遇到一些比较tricky的东西，可能需要调整fixative，固定时间，
permeabilization等等。
我推荐分离之前fix，因为那样的话可以保证... 阅读全帖 |
|
w****l
发帖数: 535 | 8 10um frozen section 用的SV2和a-bungarotoxin的double fluo stain
感觉autofluorescence很厉害，要用多少magnification看
看别人paper一对对的很漂亮
dissection，fix, section有什么讲究 |
|
g*r
发帖数: 2116 | 9 这玩意儿就是玻璃上有些凹槽 可以盛点溶液... 做个dissection/staining啥的有时候
还挺方便 实验室里很多 没想到还挺贵的 |
|
n********k
发帖数: 2818 | 10 我想我会老死在bench...
btw, I heard Andrew fire is 仍然做实验, fred gage loves to show people how
to do dissection:))) |
|
l**********1
发帖数: 5204 | 11 Alleviating interactions.
Please go to AMPK(Sip2) Nature paper:
Functional dissection of lysine deacetylases reveals that HDAC1 and p300
regulate AMPK (2012)
link:
//www.ncbi.nlm.nih.gov/pubmed/22318606
its pp251
called both as
[synthetic lethal (negative/aggravating) and rescue (positive/alleviating)
interactions]
more reference:
Acetylation of yeast AMPK controls intrinsic aging independently of caloric
Cell 146: 969-
//www.ncbi.nlm.nih.gov/pubmed/21906795 |
|
v****e
发帖数: 131 | 12 最近用老鼠组织提RNA，准备做microarray，可是RNA降解了。损失了`20个老鼠，非常
不幸。
dissect tissue，每个老鼠take 10分钟，因为比较难剥。是不是这个原因？放在RNA
LATER里比较难剥，因为变硬了。
然后就直接把tiisue放在液氮里了。抽的时候，放在trizol里。用匀浆器打散，抽提。
最后用RNAEASY纯化，
大家觉得有什么问题？有人和我说用液氮磨碎组织再加trizol，有用吗？谢谢 |
|
c****1
发帖数: 1095 | 13 好贴留名。
顺便问问，有用RNAlater dissection sample，然后做western的么？
我试过，好像没有问题。 |
|
n********k
发帖数: 2818 | 14 Any similar report before and how novel and well dissected? Big boss? tRNA/
small but non-miRNA/siRNA regulation is still very hot...If it is well
executed study and very novel, it could go as high as you could ever
imagine/like....there was a cell paper on tRNA as a potential oncogene
several ys ago but no follow up so far...I actually have noticed one miRNA-
like sncRNA of tRNA origin in my own research and considering how abundant
and extremely dramatic the change is, I suspect it would be v... 阅读全帖 |
|
l**********1
发帖数: 5204 | 15 sunnday 个把月前问 酵母内的 协同致死的叫synthetic lethal factors
而协同延命的叫啥
matrix 无意中搜到一篇nature
Lin YY et al. (2012)
Functional dissection of lysine deacetylases reveals that HDAC1 and p300
regulate AMPK.
Nature. 482: 251-5.
//www.ncbi.nlm.nih.gov/pubmed/22318606
就回帖她/他
可能叫
synthetic rescue factors
楼主你的原开贴 还有 antagonistic rescue factors 的可能性
当然 antagonistic lethal factors 的话 达尔文演化论中的不适应者 可能就无后了
具体如何定位pathway 0.44 概率以上的经典规则 不一定适用
-----just matrix two cents |
|
l**********1
发帖数: 5204 | 16 LZ can find almost answers from PhD dissertation
by Phadke, Nikhil D.
Issue Date: 2011 Univ of Michigan
"Use Of Proteomics For The Analysis Of Hard-To-Dissect Biological
Samples."
link:
//deepblue.lib.umich.edu/handle/2027.42/84528
and PDF link:
//deepblue.lib.umich.edu/bitstream/2027.42/84528/1/nphadke_1.pdf |
|
o**a
发帖数: 86 | 17 Stroke related to carotid artery dissection in a young patient with Takayasu
arteritis, systemic lupus erythematosus and antiphospholipid antibody
syndrome.
Caso V, Paciaroni M, Parnetti L, Cardaioli G, Biscarini L, Acciarini AE,
Rubino S, Gallai V.
Source
Stroke Unit, Department of Neuroscience, Unit of Internal Medicine and
Oncological Sciences, University of Perugia, Italy. |
|
y***n
发帖数: 109 | 18 养神经细胞的时候，DIV 是从哪一天算是第一天？
dissection的那一天是DIV 0 还是 1？ |
|
d*********w
发帖数: 6 | 19 这个Lin YY 很高产啊，三年内三篇CNS。高度怀疑其他paper的可信度。。。那个Lu JY
好像是他老婆。
Functional dissection of lysine deacetylases reveals that HDAC1 and p300
regulate AMPK.
Lin YY, Kiihl S, Suhail Y, Liu SY, Chou YH, Kuang Z, Lu JY, Khor CN, Lin CL,
Bader JS, Irizarry R, Boeke JD.
Nature. 2012 Feb 8;482(7384):251-5. doi: 10.1038/nature10804.
Acetylation of yeast AMPK controls intrinsic aging independently of caloric
restriction.
Lu JY, Lin YY, Sheu JC, Wu JT, Lee FJ, Chen Y, Lin MI, Chiang FT, Tai TY,
Berger SL, Zhao Y, Ts... 阅读全帖 |
|
b*****n
发帖数: 1841 | 20 说不客气点，你的认知水平在一个比较低的层次。这可能跟你老板的视野，周围同事的
水平，你专注于当前做的事情等有关，需要跟更多生物领域的老板博后博士生聊天探讨
提高。这好比是我大学毕业觉得分子生物学是了不得的工具，克隆个基因那是很
fashion的东西，酵母双杂交钓蛋白相互作用牛上天了。这些东西都是因为自己在一个
封闭落后的环境中臆想出来的。多年过去的今天，这些东西要么被淘汰（现在基本上没
有人做酵母双杂交了），要么成了非常简单不值一提的工具了。
现在做分子生物学和生物化学的，dissect pathway的基本上难发好paper难拿funding
了，混得开的也是跟那些热门的pathway靠近。
生态的如果知道分子生物学一套的，这两者都很通的人的确不是很多，那你要找到一个
着眼点，让这两者相得益彰，如果冲着这个去的，是可以学一些生化分子生物学的东西
（但是我非常怀疑你现在能有高度想到自己以后要做什么很有意义的方向；自己懵懂认
为有意义的不算）。分子植物生理方面的，要做得很好发很好的文章也是可遇不可求；
另外如前面的网友所说，生化和分子生物学这种东西太简单了，学个半年不太笨的话基
本就上手... 阅读全帖 |
|
y****o
发帖数: 26 | 21 本人女，<30岁，有美国的master, 在美国有5年实验室工作经验。因6月底要搬家到麻
省，现在正在找技术员的工作。
我有丰富的动物实验经验，including breeding, genotyping, bleeding, IP
injection, tail vein injection, retro-orbital injections, intragastric
administration, dissection of rats & mice, lethal bone marrow transplant.同
时也擅长细胞培养，包括cell line, primary cell culture, hybridoma cell
culture。常用的bench work 基本都做过，比如FACS，Immunohistochemistry， ELISA
， Production and purification of monoclonal antibody from hybridoma cells，
Western blotting, Quantitative real-time... 阅读全帖 |
|
p*****c
发帖数: 20445 | 22 A-D E-K L-P R-Z
Peter Baumann, Ph.D.
HHMI Early Career Scientist
Stowers Institute for Medical Research
Peter Baumann’s research focuses on beginnings and endings: beginnings in
the form of reproduction in unisexual lizards and endings in the form of the
telomeres that protect chromosome tips.
Each time chromosomes are copied, telomeres—specialized stretches of DNA
that extend from the ends of each chromosome—become progressively shorter.
With each cell division, they ero... 阅读全帖 |
|
p*****c
发帖数: 20445 | 23 Marc R. Freeman, Ph.D.
HHMI Early Career Scientist
University of Massachusetts Medical School
Neurons hog the attention, but our brains couldn’t operate without the
unassuming neural cells called glia. Marc Freeman is working to bring these
overlooked cells—which represent about 60 percent of brain cells—their
share of credit.
Freeman has been dissecting the role of glia in brain development, function,
and healing after injury, when glia hurry to the damaged area and devour
cellular debris to al... 阅读全帖 |
|
S******9
发帖数: 2837 | 24 第一次做，有没有视屏推荐？
或者有好的protocol？
感谢 |
|
S******9
发帖数: 2837 | 25 我想把spinal cord 取出来做切片染色 |
|
n***w
发帖数: 2405 | 26 昨天和同学聊天，他指了笔记本上western blot，有spinal cord。我很好奇的问你是
怎么取出来的。
他说先把脊柱从头到尾出来,然后用18gauge的针头从一头冲，就出来了。
你要不试试？ |
|
S******9
发帖数: 2837 | 27 我看文献那是一种方法
还有一种是从头到尾解剖出来。
你的内皮特异性的老鼠做的如何了？ |
|
n***w
发帖数: 2405 | 28 你们那里没有人做脊柱那方面的吗？我可以详细帮你问问。
等老鼠breeding要等死我。现在做一些基本的characterization，也在合作一些我们做
不了的。想秋季就把文章投出去，然后该来要老鼠的要老鼠，我们就做我们擅长的。 |
|
|
S******9
发帖数: 2837 | 30 我看了，但是这样拔去拔来的，我怕组织结构有损害。
如果不行，估计我也就这样解剖了，和以前解剖人的尸体一样 |
|
f*******a
发帖数: 671 | 31 Flushing out the spinal cord using a needle does not work for PFA fixed
tissue, but if it is fresh tissue, it works very well.
If it is PFA fixed, perform a laminectomy from sacral to cervical spinal
cord. You need to have a very fine scissor. Just remove all the lamina with
its muscle as a whole. The whole procedure takes only 1~2 minute.
Sorry that I can not type Chinese here and my English is hard to understand. |
|
S******9
发帖数: 2837 | 32 谢谢
你的英语很好，我全部理解了
有问题再随时请教。
第一次做multiple sclerosis实验，小鼠已经诱导好了，准备取标本了。希望可以成功
with
understand. |
|
h****e
发帖数: 138 | 33 “在任何学科领域，是否有一个好导师将决定学生的研究生涯初期的成败。” [1]
什么样的导师是好导师呢？2005年，Nature创立了the Nature awards for mentoring
in science，用于表彰在指导研究生方面卓有成效的导师。有关人员广泛收集总结了学
生们对其导师的评价，总结成了给导师的16条建议 [2]，如对学生的智力与能力的尊重
，对学生职业发展（甚至选择非学术道路）的尊重，等等。
下文分别列出这16条建议，并引用学生的comments作为例证。基于我与自己的博士导师
&& 我与本科期间的两个导师的各种经历，我highlight了这些comment里让我深有感触
的地方。在异国求学，一个supportive的好导师是如父亲一般的存在（撒花）~
此外，我认为导师“nice”这个说法太笼统。我认为挑导师主要有两个维度：一个是做
人，一个是治学。真正的好导师，做人方面要正直、得体，尊重学生，让学生如沐春风
，心情愉快，感到个人价值受到认可；做研究方面，要非常responsible，对学生的研
究生涯发展很给力，所以push至少比放羊好一点，让学生紧张一点总... 阅读全帖 |
|
X******n
发帖数: 914 | 34 All the authors have signed this retraction with the exception of Y.-Y.L.,
who is deceased. |
|
|
g*********3
发帖数: 177 | 36 这篇文章。。。。也能retract啊。。。LZ能否给出链接，google了下没找到出处。谢
谢。 |
|
p*****c
发帖数: 20445 | 37 这篇文章背后的故事很多。。。好像美陆台都牵涉到了？ |
|
|
|
H****N
发帖数: 997 | 40 This makes one wonder how many more papers in CNS are actually junk but will
never be found out. |
|
l**********1
发帖数: 5204 | 41 wet bio fields if WB figure used high contaction those CNS also N sister if
IF higher than
20.0 might be >1/3
there are fasle result/s. >1/10 could be retracted within one or two decade.
..
will |
|
|
A******y
发帖数: 2041 | 43 That's right, it turn out high impact paper in lower IF jounrals have best
reproducibility. If I remember correctly is above 70% by someone completely
independent.
if
decade. |
|
m******g
发帖数: 467 | 44 这期Nature好赞！
Nature 14.1.30
非常推荐：
1. Molecular biology: A second layer of information in RNA
1) In vivo genome-wide profiling of RNA secondary structure reveals novel
regulatory features
2) Genome-wide probing of RNA structure reveals active unfolding of mRNA
structures in vivo
3) Landscape and variation of RNA secondary structure across the human
transcriptome
2. Cell biology: Potency unchained
Mildly acidic conditions triggers potential for a new cell state with
diverse potentials
1) Bidirecti... 阅读全帖 |
|
m******g
发帖数: 467 | 45 这期Nature好赞！
Nature 14.1.30
非常推荐：
1. Molecular biology: A second layer of information in RNA
1) In vivo genome-wide profiling of RNA secondary structure reveals novel
regulatory features
2) Genome-wide probing of RNA structure reveals active unfolding of mRNA
structures in vivo
3) Landscape and variation of RNA secondary structure across the human
transcriptome
2. Cell biology: Potency unchained
Mildly acidic conditions triggers potential for a new cell state with
diverse potentials
1) Bidirecti... 阅读全帖 |
|
m******g
发帖数: 467 | 46 Nature 14.04.17
稍微了解一下：
1. miRNAs trigger widespread epigenetically activated siRNAs from
transposons in Arabidopsis
easiRNA resemble secondary siRNA
latent mechanism that targets tranposon transcripts
小知识：
1. Sperm RNA carries marks of trauma
Stress alters miRNA expression level in mice
which leads to depressive behavior in later generations
stress on mother, F2 generation of F1 males shows disorder
so does F3
2. Dissecting appetite
The hunger circuits has been mapped in mouse
3. Mouse already... 阅读全帖 |
|
h*******o
发帖数: 4884 | 47 Adult neuronal culture is indeed very tricky. And most of what you get is
debris.
But to dissect out cerebral cortex is easy. By 软脑膜, do you mean meninges?
You can actually start from inside. Use a sharp pair of tweezers to snip out
the meniges on the outside, then flip open the hemisphere, there you will
be able to identify hippocampus and cerebral cortex. You can use tweezers to
snip out cortical pieces.
There will be a lot of debris (actually the majority of the fresh tissue
culture will be d... 阅读全帖 |
|
D*a
发帖数: 6830 | 48 如果是看肺的话肯定是要刚死就做了，死久了看不出来什么。一般憋死半小时之内就死
了，时间再长应该不是憋死的，是也是血液之类有问题，或者是代谢问题，肌肉问题，
等等，而不可能是肺。具体细节例如
Close monitoring at birth indicated that the Igflr(-I-) dwarfs were born
alive but, despite visible efforts to breathe, became cyanotic and died
within minutes. Lung tissue dissected from several mutant neonates obtained
from independent litters failed to float on water, indicating that air never
reached the alveoli.
（ http://www.ncbi.nlm.nih.gov/pubmed/8402901）
关键是先搞明白怎么死的，这个你得全程观察。现在说太多种死法背后的原因也没什么
意... 阅读全帖 |
|
d********r
发帖数: 3279 | 49 My thoughts so far:
I think the "uniqueness" of using optogenetics in invertebrates, worms in
particular, is still essentially related to the general advantages of using
this model organism in the first place: transparent, small size, simple
nervous system and genetic screens.
For ephys analysis of synaptic function in dissected preps using
optogenetics, there is no big difference between worms and mice. For whole
animal behavioral studies under light stimulation, worms under a LED is a
lot eas... 阅读全帖 |
|
d*****s
发帖数: 647 | 50 Nature. 2013 Nov 7;503(7474):146. doi: 10.1038/nature12727.
Retraction: Functional dissection of lysine deacetylases reveals that HDAC1
and p300 regulate AMPK.
Yu-yi Lin, Samara Kiihl, Yasir Suhail, Shang-Yun Liu, Yi-hsuan Chou, Zheng
Kuang, Jin-ying Lu, Chin Ni Khor, Chi-Long Lin, Joel S. Bader, Rafael
Irizarry & Jef D. Boeke
Nature 482, 251'255 (2012); doi:10.1038/nature10804
In response to a concern raised by a reader about inconsistencies in our
Letter between the results from the primary mi... 阅读全帖 |
|
