a*******g
发帖数: 33 | 1 Hello. I work with a line of myeloid cell specific gene KO mice recombined
by LysM-Cre. Boss would like to know the KO efficiency in adipose
macrophages. In stromal vascular cells from adipose tissue, CD11b magnetic
beads were used to seperate macrophages. Surprisingly, the CD11b negative
group showed a pure monocyte/macrophage group judged from morphology in Diff
quick stained slides, while the positive population included more
neutrophils.Anyone has similar adipose tissue macrophage seperation... 阅读全帖 |
|
n***w
发帖数: 2405 | 2 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 |
|
j******2
发帖数: 544 | 3 谢谢大家回复！ 这样我就放心了。
这个Figure 1挺重要的，是唯一一个in vivo data (显示tissue morphology的变化)。
figure 2-10都是基于tissue中分离出来的cell所做的实验，都是in vitro的，所以力
量弱一些。
之前他分析的电镜图片，如果没有他，我一个人确实handle不了。不过大家这样一说，
我就放心了，我并没有占他便宜。 |
|
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o********r
发帖数: 775 | 5 http://murphylab.web.cmu.edu/PostdoctoralPositions-MurphyGroup.
Another one:
Postdoctoral Fellow Openings – Biomedical Computer Vision
Ray and Stephanie Lane Center for Computational Biology
School of Computer Science
Carnegie Mellon University
The Murphy and Rohde groups within the Lane Center for Computational Biology
, a department within the School of Computer Science at Carnegie Mellon
University, are seeking two postdoctoral fellows for a new project in the
area of computer vision for digi... 阅读全帖 |
|
K******S
发帖数: 10109 | 6 【 以下文字转载自 Joke 讨论区 】
发信人: Coho (Coho), 信区: Joke
标 题: 科学论文：粉木耳最受欢迎
发信站: BBS 未名空间站 (Sat Apr 28 16:25:56 2012, 美东)
发信人: TravisBickle (Travis), 信区: Military
标 题: 科学论文：粉木耳最受欢迎
发信站: BBS 未名空间站 (Sat Apr 28 16:22:01 2012, 美东)
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.p
Abstract
We found a relative preference for pinker genital images with redder
genitalia rated significantly less sexually attractive. This effect was
independent of raters' prior sexual experience and variation in fem... 阅读全帖 |
|
n***w
发帖数: 2405 | 7 直接用DNase做positive control.
这样可以告诉你至少两个信息。
1 你的TUNNEL工作了没？
2 阳性对照下的morphology。
上面说的对，你需要多重复重复。
不过在有tunnel信号的时候，组织不一定会mess up. |
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c********n
发帖数: 67 | 8 Simultaneous imaging of morphological plasticity and calcium dynamics in
dendrites
Thanks a lot for your kindly help
h***[email protected] |
|
w******y
发帖数: 2504 | 9 我可能没表达清楚。这个课题有两个亮点：
1。 这个基因敲除了之后，在HFD引起的肥胖症中有非常显著的心功能保护的作用，也
是我最最感兴趣的，但是不知道该怎么弄了，因为心脏MORPHOLOGY和心功能完全是正常
的。之前在其它的模型中，这个基因敲除之后心功能会变的更坏；
2。 这个基因敲除之后呢，老鼠进食（LAB CHOW 和HFD）都显著性的减少了，但是呢，
老鼠最后还是很肥，最后体重的绝对值和WT的一样。这个是这个课题中的一个有意思的
点。
我就是一搞心脏的，所以对心脏感兴趣。这个敲除了之后，心功能完全正常了，而且是
在肥胖的老鼠中，而且跟其他的模型中的作用完全相反的。 |
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l**********1
发帖数: 5204 | 10 if originally omitted below attached figure shown right pathway
then no way can bet below report:
Levy SF et al.(2012)
Bet hedging in yeast by heterogeneous, age-correlated expression of a stress
protectant.
PLoS Biol. 10: e1001325.
Abstract
Genetically identical cells grown in the same culture display striking cell-
to-cell heterogeneity in gene expression and other traits. A crucial
challenge is to understand how much of this heterogeneity reflects the noise
tolerance of a robust system and ho... 阅读全帖 |
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s****9
发帖数: 932 | 11 If they express T cell receptor (e.g. CD3 or TCRbeta), they are bona fide T
cells. You can stimulate them with plated bounded CD3/CD28 to see whether
they can proliferate, which is a function indicator for T cells.
Regarding macrophage, you could stain a few typical macrophage marker, such
as F4/80, CD11b, lysozyme M as you mentioned. The morphology of macrophage
is quite different from T cells as well.
You do not need a reference. Instead, you should add some splenic T and
macrophage as posi... 阅读全帖 |
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A******y
发帖数: 2041 | 12 Mitochondria is not round. It has network morphology, unless your cell is
dying. |
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d*****r
发帖数: 2583 | 13 It's because above the gene and molecular level, there's no established high
-throughput techniques, and also there's no well defined questions to ask.
Another reason is, above gene and molecular level, even at multi-gene and
multi-molecular level, it's a complex system. There's no analytical solution
for almost all the complex systems, which means prediction is almost
impossible. Of course you can always do numerical simulation, but that will
be impossible to generate something clean and simple... 阅读全帖 |
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m*********7
发帖数: 5207 | 14 Agree.
high
solution
will
2.
morphology: |
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m********8
发帖数: 157 | 15 following SunnyDay's suggestion, I will introduce some edible mushrooms here
in case some find it useful. I have been living in New England area for 10
years, and I know edible fungi in this area well. For mushrooms collected in
southern states and along west coast line, I am not the to-go person for
accurate identification.
I will post useful links with pictures to edible mushrooms I found common in
New England, and I will report these mushroom when I first spot them in
their growing season.
To... 阅读全帖 |
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O******e
发帖数: 4845 | 16 Press Release
2012-10-08
The Nobel Assembly at Karolinska Institutet has today decided to award
The Nobel Prize in Physiology or Medicine 2012
jointly to
John B. Gurdon and Shinya Yamanaka
for the discovery that mature cells can be reprogrammed
to become pluripotent
Summary
The Nobel Prize recognizes two scientists who discovered that mature, specia
lised cells can be reprogrammed to become immature cells capable of developi
ng into all tissues of the body. Their findings have revolutionised our... 阅读全帖 |
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m***i
发帖数: 4637 | 17 JEEM IS THE FATHER OF THE JOURNAL "DEVELOPMENT".
AND GURDON IS THE CHAIRMAN OF THE COB: COMPANY OF BIOLOGISTS
COB OWNS THE JOURNAL DEVELOPMENT, IN ADDITION TO THE JOURNAL OF CELL SCIENCE
, DMM AND "BIOLOGY OPEN" (SIMILAR TO PLOS ONE, JUST LAUNCHED) |
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w*******5
发帖数: 831 | 18 Anderson, N.O., Plattes, A., Opitz, E., Younis, A. Transgressive
segregant, interspecific hybrids between lilium ×formolongi and L. martagon
with unique morphology ( Conference Paper ) . Acta Horticulturae 2011, 900
请发到w**********[email protected]，谢谢！ |
|
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e***l
发帖数: 33 | 20 Company Introduction
SOTIO is a European biotechnology company developing new therapies based on
activated dendritic cells, focusing on the treatment of cancer and
autoimmune diseases. Our mission is to develop new medical therapies using
our proprietary cell-based technologies to treat highly unmet medical
conditions using our immunotherapy platform. Sotio (Beijing), a fast growing
subsidiary focusing on China market, is looking for talented, motivated,
dynamic employees to join its internation... 阅读全帖 |
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d**p
发帖数: 149 | 21 I know the small GTPases involves. |
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s******s
发帖数: 13035 | 22 不知道你说的变化有多特殊。一般under stress或者cell cycle
有点变化都会体现在Morphalogy上，会有无数gene |
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w***a
发帖数: 1053 | 23 Actin remodeling,cell adhesion
太多基因了
让sunnyday来说说 |
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w***a
发帖数: 4361 | 24 做了啥treat，用的啥细胞都不说，那就木法猜。 |
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m********m
发帖数: 263 | 25 不好意思，我用的是HepG2 cells 和MCF-7， knockdown一个激酶后，cells变得很“邪
恶，张牙舞爪”的样子 |
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m********m
发帖数: 263 | 26 Thanks! this is very helpful |
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Z******5
发帖数: 435 | 27 建议检测一下differentiation marker和EMT marker |
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a****i
发帖数: 21 | 28 Scale: a chemical approach for fluorescence imaging and reconstruction of
transparent mouse brain
Nature Neuroscience 14, 1481–1488 (2011) doi:10.1038/nn.2928
版上牛人能否解释下，和这篇文章比，这片nature article强在哪儿？
karl太cool了，不光在brain的activity领域开创Optogenetics， 如果这篇文章大规模
应用，在brain morphology领域贡献也很大。当年Cajal就凭借Golgi staining，开创
现代神经生物学。。。 |
|
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p*****c
发帖数: 20445 | 30 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 |
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e*******r
发帖数: 275 | 31 请问版上哪位可不可以refer我一些审稿机会，最近心力交瘁，急需申请绿卡。先谢过
了。
方向大致是dendritic spines, dendritic spine morphology,actin, cytoskeleton,
receptor tyrosine kinase, opioid receptors, GPCR, functional selectivity,
arrestin recruitment. etc.
a**********[email protected] |
|
m******g
发帖数: 467 | 32 Nature 14.02.27
推荐：
1. Genetics of single-cell protein abundance variation in large yeast
populations
稍微了解一下：
1. Sequential evolution of bacterial morphology by co-option of a
developmental regulator
小知识：
1. Molecule makes blind mice see light
2. Monkey brains wired to share
prisoner's dilemma
3. Crowd-sourcing: Strength in numbers
4. Skin pigmentation provides evidence of convergent melanism in extinct
marine
reptiles
5. A discrete genetic locus confers xyloglucan metabolism in select human
gu... 阅读全帖 |
|
m******g
发帖数: 467 | 33 PNAS 14.04.22
推荐：
1. DNA methylation and evolution of duplicate genes
DNA methylation clearly distinguishes young and old duplicates
DNA methylation of duplicate genes are associated with functional
differences in expression
[wish to see some experimental evolution result in the future]
稍微了解一下：
1. Massive programmed translational jumping in mitochondria
Report 81 translational bypassing elements (byps) in mitochondria of the
yeast Magnusiomyces capitatus
Byp-like sequences lack bypassing activi... 阅读全帖 |
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l*******i
发帖数: 153 | 34 Xist下调是RNA-seq的结果，还是qPCR结果？这一点比较奇怪，与X chromosome
inactivation不一致。
按理说，在mESC中，X chromosome是active的，即XaXa，Xist应该不表达才对。Xist表
达时，会把其中一条X chromosome“包裹”起来，这条chromosome进而失活。X
inactivation一般指发生在female mEpiSC与分化的细胞中.
你这个结果与理论相反，X chromosome基因下调时，Xist竟然也降低。Xist在对照和
knockdown细胞中的表达分别是多少(RPKM or CT value)？
另一个问题，由于你用的培养条件是serum+LIF,这会导致heterogeneity.建议你用2i/
LIF.
knockdown mESC分化明显吗？ colony morphology改变如何，还是依旧维持与WT一样的
形态？
如果你确定用的是female mESCs的话，建议先做H3K27me3 staining,看看X chromosome
是不是失活的。 |
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i*********0
发帖数: 915 | 35
是RNA-seq的结果。我也觉得很奇怪，
这一点比较奇怪，与X chromosome
Xist在对照和
这个我要去问一下我们的搞生物信息学的。
ok！
分化不明显。
colony morphology改变如何，还是依旧维持与WT一样的
colony looks like the control shRNA clone。 shiny and clear boundary |
|
p**t
发帖数: 160 | 36 KO mice?
You should also check other myeloid cell populations, e.g. DC, MDSC, even
CDP, MDP. Absolute numbers and percentages.
As for macrophage, did you check F4/80, MerTk, MHCII, CD40, CD80, CD86, PD-
L1, PD-L2, morphology, and phagocytosis?
And, the phenotypic change is generally on all lymphoid organ and tissue
macrophage subsets? |
|
m******g
发帖数: 467 | 37 PNAS 14.10.07
稍微了解一下：
1. Alternating antibiotic treatments constrain evolutionary paths to
multidrug resistance
小知识：
1. Structural adaptations to diverse fighting styles in sexually selected
weapons
horns morphology in rhinoceros beetle species |
|
l*******i
发帖数: 153 | 38 照着Cell Stem Cell与Stell Cell Report的Editorial Board找就行了。stem cell的
牛人基本上都在这两个Board里了。
另外，干细胞，特别死胚胎干细胞与iPSC领域内的问题，很大一部分不是由于人为造假
，而是由于干细胞的特性决定的。与其他的细胞不一样，干细胞对enviroment的适应性
太差，很容易就会发生改变，而且很多时候，仅凭少数的评价指标（比如morphology,
marker gene expression等）根本就区分不出来。
但不可否认的是，干细胞确实是一个非常好的modeling platform；最近几年，生物与
基础医学领域的许多大的突破，都得益于干细胞的研究，比如epigenetics, NGS的应用
，translation medicine的发展，等等。 |
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e*r
发帖数: 103 | 39 A contribution to the morphology and life history of Aristolochia
多谢！ |
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b**********a
发帖数: 930 | 40 楼主说的对，现在的化合物库和化合物设计、合成、in vitro筛选等都不是瓶颈，in
vitro筛选的效力惊人，只要有足够的化合物，in vitro活性都可以得到。
真正的瓶颈在in vivo筛选，这里面的最大问题的动物模型，现在最常用的是大鼠和小
鼠。这些动物in vivo和人类的相关性、预测性等完全没有办法，动物模型本身的病理
状态和人疾病的相关性等。如果哪一天猴子模型可以很普遍的应用，可能这些预测性和
相关性会好很多。
《PNAS》最近提到的生命科学类11月最受关注的十篇论文中提到小鼠模型须小心使用，http://www.biomart.cn/news/103/129225.htm
小鼠被广泛用于模拟人类代谢、疾病和药物应答，是医学研究中的一个基本工具。然而
斯坦福大学的研究指出，人类和小鼠的基因表达存在着惊人的差异，不论是蛋白编码基
因还是非编码基因。这项研究发表在十一月十七日的美国国家科学院院刊PNAS杂志上。
相信在未来的不久对大鼠基因表达也会有类似的研究，这样应该可以比较好的解释为什
么动物模型的相关性和预测性是如此之差。
小鼠被广泛用于模拟人类代谢、疾病和药物应答，是医... 阅读全帖 |
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n*******l
发帖数: 190 | 41 http://www.ncbi.nlm.nih.gov/pubmed/25700174
Science. 2015 Feb 19. pii: aaa1934. [Epub ahead of print]
Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-
seq.
Zeisel A1, Manchado AB1, Codeluppi S1, Lönnerberg P1, La Manno G1, Jur
éus A1, Marques S1, Munguba H1, He L2, Betsholtz C3, Rolny C4, Castelo-
Branco G1, Hjerling-Leffler J5, Linnarsson S5.
Author information
Abstract
The mammalian cerebral cortex supports cognitive functions such as
sensorimotor integration, m... 阅读全帖 |
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C*****8
发帖数: 102 | 42 International PhD Scholarship in Microbial Cytobiology
The Imagine initiative at the University of Sheffield is an exciting venture
to combine cutting edge ultra high-resolution light microscopy, cryo-
electron microscopy and biochemistry to answer fundamental questions in
bacterial ultrastructure and function. An international PhD studentship [
open to UK, EU and international applicants] is available in Dr. Egbert
Hoiczyk’s lab that focuses on newly discovered bacterial cytoskeletal
proteins a... 阅读全帖 |
|
发帖数: 1 | 43 Postdoctoral Positions Available
Two postdoctoral positions are immediately available for studies in
neurophysiology focusing on the role of voltage-gated potassium channels in
regulating hippocampal circuit development and function. The candidate will
be working with a team of highly interactive electrophysiologists and
molecular biologists at the Eunice Kennedy Shriver National Institutes of
Child Health and Human Development (NICHD), National Institues of Health (
NIH).
Skills and Qualifica... 阅读全帖 |
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c********n
发帖数: 225 | 44 第四类“重复” 验证实验信息：实名, 网络、论坛发布
- 重复、拓展验证实验不能支持NgAgo基因组编辑功能
- 包含有具体实验信息（eg实验材料，步骤，etc）以及发布方具体信
息的发布报告
共计13则
细节请参见
Date：2016年10月10日
Title：13位科学家实名呼吁对韩春雨启动调查：为了中国学界的名声
Link：
http://www.thepaper.cn/newsDetail_forward_1541143_1
增加实验细节请参见：
Date：2016年10月11日
Title：【赛先生】13个课题组实名声明：无法重复韩春雨实验
Link:
http://chuansong.me/n/943170546756
其中， 根据一些实验室数据推论，“NgAgo或许不具备双链DNA切割活性”
“北京大学生物动态光学成像中心研究员孙育杰的实验室专长荧光成像。。。。孙育杰
进一步解释：端粒有一个特点，它有数千个重复片段，如果用CRISPR或者TALEN这样的
技术去target（靶向）端粒的重复片段的话，由于它有上千次重复，数百、上千个
CRISPR或TALE会富... 阅读全帖 |
|
c********n
发帖数: 225 | 45 第四类“重复” 验证实验信息：实名, 网络、论坛发布
- 重复、拓展验证实验不能支持NgAgo基因组编辑功能
- 包含有具体实验信息（eg实验材料，步骤，etc）以及发布方具体信
息的发布报告
共计13则
细节请参见
Date：2016年10月10日
Title：13位科学家实名呼吁对韩春雨启动调查：为了中国学界的名声
Link：
http://www.thepaper.cn/newsDetail_forward_1541143_1
增加实验细节请参见：
Date：2016年10月11日
Title：【赛先生】13个课题组实名声明：无法重复韩春雨实验
Link:
http://chuansong.me/n/943170546756
其中， 根据一些实验室数据推论，“NgAgo或许不具备双链DNA切割活性”
“北京大学生物动态光学成像中心研究员孙育杰的实验室专长荧光成像。。。。孙育杰
进一步解释：端粒有一个特点，它有数千个重复片段，如果用CRISPR或者TALEN这样的
技术去target（靶向）端粒的重复片段的话，由于它有上千次重复，数百、上千个
CRISPR或TALE会富... 阅读全帖 |
|
c********n
发帖数: 225 | 46 对于韩春雨NBT文章无法重复的问题
需要从Figure 1 开始一步步看
作者的contributions
1. C.H. conceived the study and designed the experiments.
文章 “Story” “Idea” 的设计与发起人
2. X.Z.S. provided intellectual advice on the project and experimental
design.
实验设计， “One guide-faithful” -Figure 2d？
3. C.H. performed mammalian genome editing.
（Figure 4,5）
文章里让科研界最感兴趣的结果，也是无法被重复验证的主要问题
4. F.G. performed the BLAST search and the in vitro cleavage experiments.
（Figure 1, supplementary figure 2）
大肠杆菌E coli 表达的蛋白
NgAgo 基因编辑功能的初步 in vitro 数据支持
注... 阅读全帖 |
|
c********n
发帖数: 225 | 47 【再贴一次】
对于韩春雨NBT文章无法重复的问题
需要从Figure 1 开始一步步看
作者的contributions
1. C.H. conceived the study and designed the experiments.
文章 “Story” “Idea” 的设计与发起人
2. X.Z.S. provided intellectual advice on the project and experimental
design.
实验设计， “One guide-faithful” -Figure 2d？
3. C.H. performed mammalian genome editing.
（Figure 4,5）
文章里让科研界最感兴趣的结果，也是无法被重复验证的主要问题
4. F.G. performed the BLAST search and the in vitro cleavage experiments.
（Figure 1, Figure 2？，Figure 3a，supplementary figure 2）
大肠杆菌E coli 表达的蛋白
NgAg... 阅读全帖 |
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