c******6
发帖数: 928 | 1 C12 - Resazurin viability assay.
O'Brien J, Wilson I, Orton T, Pognan F. Investigation of the Alamar Blue (
resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity
. European Journal of Biochemistry. 2000;267:5421-6. |
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h*******o
发帖数: 4884 | 2 3-NPA does that. 3-NPA directly inhibits succinate dehydrogenase activity.
At sub-toxic doses cells are still viable, but SDH activity could be significantly inhibited.
MTT/MTS/Calcein Am/Resazurin assays are often over-used or somewhat used
incorrectly to measure cell viability/cell number.
your
seen |
|
h*******o
发帖数: 4884 | 3 As mentioned above, a lot of the cytotoxicity/cell viability assays
are based on mitochondrial function, particularly SDH activity. This renders
a lot of the assays under question when the "manipulation/treatment" have
potential impacts on mitochondrial function.
For cancer cell lines, it is less of an issue because of the
notorious "warburg" effect through which cancer cells are more glycolytic
and rely much less on mitochondrial OXPHOS.
But for primary cultures, like... 阅读全帖 |
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k***g
发帖数: 4904 | 4 试了试，细胞太小，显微镜下没法判断是什么颜色啊、
有人能说说不，两三微米大的细菌，能用Trypan Blue测活动性么？ |
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W****C
发帖数: 1937 | 5 主要是怎么提高细胞的viability，分离过程（我用MACS去除其他细胞)有哪些trick？
激活的方法比较多， LPS, IgM, IL4, PMA...怎么激活最好？ |
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D*a
发帖数: 6830 | 6 看了看那篇cell。。。
这个，他们说，“Fitness” in this context is similar to that measured in
yeast in either liquid growth or colony growth assays and is an aggregate
property of the population; a defect in fitness could result from many
different causes from changes in brood size, viability, or growth rate, or
other more subtle defects in organism function. (result 第一段末尾)
下面更简化成了fitness = number of animals /remaining food (bacteria)
这个首先存疑。本来在实验室里养了千秋万代的虫子，当然是适应了实验室环境，然后
敲掉一些genes，发现在实验室环境中敲掉的那些基因的虫子比原来的长的慢 。... 阅读全帖 |
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w***r
发帖数: 709 | 8 No mechanistic insight right now, only detailed domain characterization.
I am not in plant field, but I don't think you could publish in journal IF>6
at the moment.
But I do think the question about evolution is interesting.
I hope this paper is helpful.
Science. 2010 Dec 17;330(6011):1682-5.
New genes in Drosophila quickly become essential.
Chen S, Zhang YE, Long M.
Abstract
To investigate the origin and evolution of essential genes, we identified
and phenotyped 195 young protein-coding genes, ... 阅读全帖 |
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d****d
发帖数: 214 | 9 I have not used Neon for a while, but I think 50% survival rate would be
reasonable to expect. Of course you'll need to optimize the voltage,
duration time, and number of pulses.
Our neighbor lab and I changed the tips and buffers ten times less
frequently than suggested by Invitrogen and it still works well. I guess the
company is counting on the consumables for hefty profits. Also we requested
a lot more (free) buffers at the time we purchased the machine.
The main advantage for electroporatio... 阅读全帖 |
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s******a
发帖数: 472 | 10 Same program works fine for small plasmid, but viability decrease
dramatically if I try to electroporate large DNA construct. Why?
cells |
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c**m
发帖数: 757 | 11 这个才是她能被MIT ＭＢＡ录取的真正原因
一个PhD没有任何管理经验，连UCLA ＭＢＡ都进不去
President, Co-founder
Advanced Degree Consulting Club at UCLA
April 2010 – February 2012 (1 year 11 months)
• Created a community of future consultants and business leaders
with advanced degrees and provide a platform for information exchange and
experience sharing.
• Led the team and generated substantial revenue by undertaking
consulting projects with local consulting firms.
• Bridge the gap between academia and consulti... 阅读全帖 |
|
n****n
发帖数: 434 | 12 = or >90% FBS will maintain the viability for a while. But of course some
cell lines are more fragile than others. |
|
A****w
发帖数: 244 | 13 最近在做baculovirus transfection一直做不出来
用了两种不同的方法，一个是BD BaculoGold DNA ＋ pVL1393-DNA.
就是follow manual
另一是Bac to Bac system， 用cellfectin II reagent。
用的是Sf-21 conditioned in Sf-900II media （大于95％viability)
怎么做都在5天transfection后看不到病态细胞
然后amplification后也没有～
请大牛指教！
感谢！ |
|
h******y
发帖数: 351 | 14 常用的MTT，XTT，Cell-Titer-Glo，LDH都跟mitochondria活性有关。如果不想用这些的
话，可以试试physical的方法，例如impedance（ACEA xCELLigence）或者confluence
（I
ncuCyte）。或者染DNA，Fluorescence-based LIVE/DEAD cell viability assays fro
m Invitrogen. |
|
s******a
发帖数: 472 | 15 研究的inhibitor用DMSO溶解，所以需要DMSO的对照。但是和没有任何处理的细胞相比
，DMSO降低基因A的表达（RT-PCR). 做了一个２个小时的time course，有一个随着
DMSO处理时间增加表达下降的趋势（和无处理组相比）。但是我使用的DMSO浓度远低于
０.５（v/v），据文献报道这个浓度该细胞不应有何影响。细胞在DMSO２个小时后
viability，形态都正常。
另外研究的基因B则不受DMSO影响(RT-PCR）。
DMSO影响基因表达的可能机制都有什么？
欢迎大家指教！多谢！！ |
|
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k***g
发帖数: 4904 | 17 我的实验只要求染活细菌就够了，不需要把死细菌有多少测出来，有没有什么荧光dye
能只染活细菌，不染死细菌？还是说只有用live/dead viability kit才可以知道有没
有活细菌？如果只能用kit，哪位大牛能给推荐个便宜点的吗，怎么染细菌的我看都400
刀以上。。 |
|
d****i
发帖数: 2346 | 18 The Bacteria Live/Dead Staining Kit enables a two-color fluorescent staining
on both live bacteria (green) and dead bacteria (red) using the two nucleic
acid dyes DMAO and EthD-III. DMAO is a green-fluorescent dye which stains
both live and dead bacteria with intact and damaged cell membranes while the
red-fluorescing EthD-III only stains dead bacteria with damaged cell
membranes. With an appropriate mixture of both dyes, viable bacteria with
intact cell membranes are stained green, whereas bac... 阅读全帖 |
|
g***j
发帖数: 40861 | 19 Penetration of Escherichia coli O157:H7 into lettuce tissues as affected by
inoculum size and temperature and the effect of chlorine treatment on cell
viability
s*****[email protected]
多谢 |
|
O******e
发帖数: 4845 | 20 我推荐大家都读一下这篇文章：
http://www.pnas.org/content/111/16/5773.full
全文：
By Bruce Alberts, Marc W. Kirschner, Shirley Tilghman, and Harold Varmus
By many measures, the biological and medical sciences are in a golden age.
That fact, which we celebrate, makes it all the more difficult to
acknowledge that the current system contains systemic flaws that are
threatening its future. A central flaw is the long-held assumption that the
enterprise will constantly expand. As a result, there is now a severe
imbalan... 阅读全帖 |
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A******y
发帖数: 2041 | 21 Btw, I made a mistake, the name should be EC50 (effective concentration).
LD (lethal dose) is usually used in vivo for animal studies.
This is how my lab does EC50: We do 12 point titration either (1/2 or 1/3)
dilution per dilution. For example, for single digit microM compounds, we
do 200 micorM, 100 micorM, 50 microM, for 12 (Log[concentration] will be
your X-axis) points...After that we determine the viability of the cells (Y
axis). Then, you use a non-linear fit program (Prism is the mos... 阅读全帖 |
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s*****g
发帖数: 7857 | 22 我做了浓度梯度3天测试。找出毒性小的最高浓度。做了4 天的处理。
24h DOX Induction, then 3 days Drug treatment with DOX.
至于蛋白量需要WB 检测。但是MTS显示了我预期的结果。显微镜下也看到了细胞形状的
变化(Apoptosis/cell death)。
Negative control and shRNA knockdown 在细胞viability上有着显著的差异！
万分感谢！ |
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r*****i
发帖数: 117 | 23 should be in the same mouse. Otherwise, how can you exclude the viability
among mice. |
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y*******g
发帖数: 4 | 24 Postdoctoral Research Fellow Position in Institute of Glycomics, Griffith
University, Gold Coast, Australia
Prof Yaoqi Zhou’s group http://sparks-lab.org is built up a strong term for his molecular biology laboratory to interact with his computational laboratory at the Institute for Glycomics, Griffith University, Gold Coast, Australia. Initial exciting discovery in protein expression optimisation, antibiotic inhibitor discovery and protein design leads to the current expansion of the team. We... 阅读全帖 |
|
发帖数: 1 | 25 【 以下文字转载自 JobMarket 讨论区 】
发信人: JuneEight (), 信区: JobMarket
标 题: 【波士顿】全职技术员/博后-药物研发相关的小鼠实验
发信站: BBS 未名空间站 (Mon May 9 14:25:30 2016, 美东)
主要工作是小鼠脑中接种肿瘤细胞及后续的监控、监测。Title可以是技术员，也可以
是博后。
本项目是与药企合作项目，对于申请者之后找工业界的工作会有帮助。
项目启动时间紧迫，所以优先当地的申请者，且只考虑有相关经验的申请者。
工作单位：Brigham and Women's Hospital
工作地点：Longwood medical area。
有意者请直接通过BWH申请，申请链接：
https://partners.taleo.net/careersection/jobdetail.ftl?job=3017
Vy0Utcf5HhI.gmail
详细说明：
Job Description
TECHNICAL RESEARCH ASSISTANT II / 40 HOURS / DAY / BWH - NEUROLO... 阅读全帖 |
|
发帖数: 1 | 26 Genome-editing
下午4:50（6 分钟前）
其他收件人： h********[email protected], c******[email protected], o*****[email protected]
将帖子翻译为中文
- 显示引用文字 -
hello guys,
we tried NgAgo plasmid cotransfection with 5'phospho-ssDNA guides and I must
say cell mortality rates are very high (toxic). We got 5'phospo-ssDNA
guides synthesised from IDT, desalt purification. Does anybody have tested
desalted verus PAGE purified ssOligos and looked for cell viability.
cheers! |
|
发帖数: 1 | 27 新发现丨细胞常温运输的高效简易方法！
2017-06-03 邓文生 等 细胞
2017年4月18日，武汉科技大学生命科学院邓文生教授在美国公共科学图书馆的期刊上
发表一篇题为“The analysis of viability for mammalian cells treated at
different temperatures and its application in cell shipment”的研究论文。主
要验证了在室温下运输细胞的简易方法的可行性。
目前，传统的细胞运输方法包括冷冻储存运输和充液法。细胞存活率较高，但是都只
能短距离运输，花费高。近几年，也还有一些关于细胞运输方法改进的报道，例如有用
agarose胶或matrigel基质与细胞混匀后，再在室温运输。这种种方法都非常繁琐复杂
，影响因素较多。
邓教授在英国做实验时，一个小的失误，意外发现室温下放置的细胞在第二天仍能保持
较高的活性。就想是否能够直接在室温的条件下运输细胞，从而简化细胞的运输方法。
为了验证这个方案的可行性，该实验室采用其常用的细胞系M2进行一系列验证。首先确
定细胞密度为1*105 ... 阅读全帖 |
|
d****i
发帖数: 360 | 28 Viability and differentiation of neural precursors on hyaluronic acid
hydrogel scaffold
Pan, L., Ren, Y., Cui, F., Xu, Q. 2009 Journal of Neuroscience Research 87
(14), pp. 3207-3220
e-mail: x**********[email protected] |
|
j******0
发帖数: 258 | 29 "Assessment of embryo viability in assisted reproductive technology:
shortcomings of current approaches and the emerging role of metabolomics"
Current opinion in obstetrics and gynecology [1040-872X] Bromer
yr:2008
vol:20 iss:3
pg:234 -241 |
|
W*****s
发帖数: 684 | 30 Carboxymethylchitosan/Poly(amidoamine) Dendrimer Nanoparticles in Central
Nervous Systems-Regenerative Medicine: Effects on Neuron/Glial Cell
Viability and Internalization Efficiency Salgado, A.J., Oliveira, J.M.,
Pirraco, R.P., Pereira, V.H., Fraga, J.S., Marques, A.P., Neves, N.M., (...)
, Sousa, N. 2010 Macromolecular Bioscience 10 (10), pp. 1130-1140
Thanks a lot! |
|
G****n
发帖数: 145 | 31 Justin Lin of CCER
[BOOK] The China miracle: Development strategy and economic reform[PDF] from
socialsecurity.orgJY Lin, F Cai… - 2003 - books.google.com
The Hong Kong Centre for Economic Research The Centre was set up as a
research and educational
trust under a trust deed signed in 1987. It supports research and publishes
studies on public
policy issues to promote understanding of economic affairs and provide
alternative policy ...
Cited by 462 - Related articles - Library Search - All 18 vers... 阅读全帖 |
|
W***n
发帖数: 11530 | 32
Areas of near-term commercial viability e.g. I'm not interested in cold fusion (remember that BYU scandal, hahaha) ,or hot fusion for that matter. |
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S*******p
发帖数: 261 | 33 Please contact Shaz Samual directly!
FYI:
One of our clients is the global leading technology provider to the
international energy industry with the world's most advanced intelligent
metering, data collection and utility software solutions. It is their vision
to provide responsible management and conservation of the world's most
valuable resources requiring optimization of the delivery and use of energy
and water. Their Systems Engineering group is rapidly expanding and we need
high impact indi... 阅读全帖 |
|
j****g
发帖数: 406 | 34 谢谢老大的建议，下面是introduction的第一段。版面有限估计贴不了更多的了，如果
老大，或者是其他哪一位有兴趣的话我可以发到邮箱里。谢谢。
The balance between protein synthesis and degradation is important for
normal function and survival of cells. While the ubiquitin-proteasome
pathway is responsible for specific clearance of short-lived proteins, long-
lived proteins are delivered to lysosomes by autophagy, which is the major
inducible pathway for general turnover of bulk cytoplasmic components
including entire organelles such as mitochondria (Klionsky and ... 阅读全帖 |
|
a**********m
发帖数: 2098 | 35 hope you could find a patent agent/attorney, or someone would contact you
but be prepared to throw in thousands of dollars before you could even see t
he viability of your patent, in case you can get one |
|
a**********m
发帖数: 2098 | 36 hope you could find a patent agent/attorney, or someone would contact you
but be prepared to throw in thousands of dollars before you could even see t
he viability of your patent, in case you can get one |
|
a*********n
发帖数: 602 | 37 What is private equity?" was the common question asked in China back in the
1990s.
The terminology was not as widespread as it is today, with only a handful of
private equity firms active in the market. I recall having to conduct a
small
seminar with a company's management team, explaining the virtues of private
equity
before any transaction terms could be discussed. Times have changed since
the
early years as China is now home to over 200 China-dedicated funds raising
over
$15bn (Rmb 103bn, 0.8... 阅读全帖 |
|
x****m
发帖数: 13 | 38 求救！！！
我的末次月经是5月28号，上周五，也就是8天前，我去做第一次阴超B超，医生给出的
结果附在后面。我是在我们的校医院看的，一直是一个MIDWIFE接待我，我也不知道美
国的这个MIDWIFE都是啥职业，感觉就跟医生一样，给我开处方，接待我，甚至给我下
诊断，我一直没有见到真正的医生。B超结果的当天我就联系她，然后她就给我发信息
说我是BLIGHTED OVUM,让我当天再去查个血，然后约时间去见她。这周的周一一早去见
她，她说我的B超当天做的血检的hCG 是四万多的样子，我也没听清具体的数字，然后
说我肯定是BLIGHTED OVUM，因为基于这样的高的荷尔蒙的浓度还没有见到胎心和胚胎
组织，她说她可以几乎肯定是不好的胚胎。说我唯一能做的就是等着自然流产，我问大
概需要等多久啊，她说她说不一定，可能在一个月之内吧。然后就想让我回家等着了。
我要求再做一次B超，然后她答应下周五再做一次B超和查血，然后就让我回家等着了。
这些天一直在网上看关于胎停育的，觉得很可怕啊。从怀孕至今一直没有过流血，甚至
深色白带都没有，身体一直很好，早孕反应也很轻，从来没有正儿八经的吐过，尤其是
这两... 阅读全帖 |
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n*******c
发帖数: 501 | 39 要回答你这些问题，光有CT和MRI是不太够的。
你父亲没有心绞痛的症状，所以主要的考虑是选择可以减少死亡率的治疗（有些病人因
为心绞痛发作药物控制不理想，所以会更加倾向于手术减少症状）
不管是否手术，你父亲都要开始药物治疗，积极控制危险因素的，其中用药有无副作用
也是是否手术的重要考量。
然后就是心功能的测试，一方面了解以前的心梗对心脏造成了多大的损害。另一方面了
解myocardial viability（PET，thallium perfusion，MRI，ECHO），这个就说有多少
还没死的心肌细胞能通过手术救活，这也是掌握手术时机的一个要点。
无论考虑上支架或搭桥，术前都是需要做血管造影评估血管及循环的真实情况，来帮助
下一步决定。
个人认为这个情况不适合支架，至于是否手术还是纯药物治疗要看其他检查结果。
如果俺是你的话，
先让父亲做一个冠脉造影，以及以上其他检查（都是无创的），然后再找能找到的最好
的心脏科专家包括心外科医生给意见，然后和你父亲讨论一下，这时候，病人自己的意
向也很重要。
另外需要考虑的是，如果决定手术，医生的技能也很重要，有时候也取决于你能找到什么样的外科医... 阅读全帖 |
|
M***D
发帖数: 249 | 40 I have 10 years of research experience in cell culture study (6 years for my
PhD and 4 years as a post-doctoral research fellow), I hope I can help you
to understand Dr. Chen's study better:
(1) Cell cultures
-Cultured normal cells have limited growth in a petri-dish due to lateral
inhibition, while cultured cancer cells grow much faster comparing to
cultured normal cells. Therefore, you must "pass" the cultured cancer cells
from one petri-dish to 2 petri-dishes more frequently in order to keep ... 阅读全帖 |
|
f******t
发帖数: 7283 | 41 上个月，GM为了那个Viability Plan，已经把High Performance Vehicle Operations
team给解散了。之后不再会有新一代的SS或者CTS-V了。 |
|
p********i
发帖数: 12365 | 42 我也听说CTS-V， impala SS, cobalt SS将没有了
可惜啊
CTS-V, 0-60MPH 3.9sec
上个月，GM为了那个Viability Plan，已经把High Performance Vehicle Operations
team给解散了。之后不再会有新一代的SS或者CTS-V了。 |
|
h******i
发帖数: 643 | 43 不要轻易忽视santa rally的规律，
前几天参加一个朋友小孩的birthday party, 有一个小孩的父亲是hedge fund manager
,和他聊了一会天，他提道近期viability比较大，这在股票市场来说其实不是一件坏事
，欧洲危机到底会对美国有多大的影响？他们的看法是最终结果不必太悲观，希腊规模
比较小，意大利主要债主在国内，回旋余地比较大，还有很多的黄金储备。从总体上来
看，欧元区贸易是平衡的，只是内部失衡。
股市需要调整，但是不必太看空，超跌的时候还是要敢于出手 |
|
s**********n
发帖数: 868 | 44 They are not directly related in any sense, but crude price probably affects
market sentiment on solar.
The viability of solar PV is more related to coal and natural gas price,
because they are widely used to produce electricity. |
|
y*****l
发帖数: 5997 | 45 【 以下文字转载自 bluechips 俱乐部 】
发信人: yaokarl (大象,面壁中), 信区: bluechips
标 题: CAT TEN
发信站: BBS 未名空间站 (Mon Feb 8 11:20:57 2010, 美东)
TEN is the biggest automotive emission control supplier for CAT and many
auto companies in Detroit area. Very good investment for the long term.
Caterpillar Inc. Joins Industry and Government Partnership to Deliver State-
of-the-Art Clean-Coal Technology
Monday 02/08/2010 9:00 AM ET - Pr Newswire
Caterpillar Inc. (NYSE: CAT) today announced its intent to join the
FutureGen A... 阅读全帖 |
|
v**********m
发帖数: 5516 | 46 Nuclear Power Has a Bright Future: Amir Adnani
The Energy Report | Jun. 5, 2012, 4:00 AM | 0 |
inShare
A A A
The Energy Report
URL
The Energy Report is a free newsletter featuring investment coverage on
fossil, renewable and alternative energies
Recent Posts
Biggest Value for Agricultural Investors Revealed: Steve Hansen
Biggest Value for Agricultural Investors Revealed: Steve Hansen
Canadian Oil Explorers Pump Profits Abroad: Frederick Kozak
RSS Feed
The Energy... 阅读全帖 |
|
v**********m
发帖数: 5516 | 47 http://seekingalpha.com/article/894061-delcath-systems-an-exclu
Delcath Systems: An Exclusive Interview With ROTH Capital Senior Analyst
September 28, 2012 | about: DCTH
Disclosure: I am long DCTH. (More...)
Today I have the pleasure of publishing an exclusive interview that took
place this week between me and one of the leading senior analysts for
medical technology, Mr. Matt Dolan of ROTH Capital Partners. Mr. Dolan has
followed the industry for nearly a decade and has been the senior analys... 阅读全帖 |
|
Q*K
发帖数: 3464 | 48 http://micello.com/blog/lbs-goes-indoors-maps-apps-and-positioning
LBS goes indoors: Maps, Apps and Positioning
December 10, 2010
WCA, Silicon Valley's Wireless Communication Alliance, conducted a panel
discussion on indoor LBS at the Qualcomm premises in Santa Clara on Thursday
, December 9th. The event was very well attended with 100+ paid attendees.
Raj Singh did an excellent job in moderating the panel by asking all the
right questions. The panelists included Ankit Agarwal of Micello, Jerry ... 阅读全帖 |
|
Q*K
发帖数: 3464 | 49 WCA, Silicon Valley's Wireless Communication Alliance, conducted a panel
discussion on indoor LBS at the Qualcomm premises in Santa Clara on Thursday
, December 9th. The event was very well attended with 100+ paid attendees.
Raj Singh did an excellent job in moderating the panel by asking all the
right questions. The panelists included Ankit Agarwal of Micello, Jerry Luk
of Presdo, Jeremy Geiger of Retailigence, Robert Schoenfield of Polaris
Wireless, Kiyo Kubo of Spotlight Mobile, Tristian Lacr... 阅读全帖 |
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