荧光下的colocalization 是不是意味着contact以及interaction?# Biology - 生物学
g*i
1 楼
小孩子的理解能力是随年龄的增长而增长的， do not let a boy do men's jobs.
F*Q
2 楼
外行向各位高手请教，多谢！
l*s
3 楼
同意
有一同学的孩子，单身母亲，特希望儿子成熟，象男人一样
总是按她的期望来说教儿子
现在13岁了，好好的儿子行为不大人不小孩的，感觉比一般孩子有点特殊
孩子就是孩子，最无忧无虑的年龄
做大人有啥好，以后有的是时间
把每个年龄段过好了才是最重要的
有一同学的孩子，单身母亲，特希望儿子成熟，象男人一样
总是按她的期望来说教儿子
现在13岁了，好好的儿子行为不大人不小孩的，感觉比一般孩子有点特殊
孩子就是孩子，最无忧无虑的年龄
做大人有啥好，以后有的是时间
把每个年龄段过好了才是最重要的
b*s
4 楼
很多co-localization只是同是定位于胞内的compartments，并不能说明两个蛋白相互
接触和作用。
比如一个vesicle上可以有很多蛋白，但他们不一定要直接接触。
要直接证明，还是需要其他手段，live imaging可以用， FRET或者BIFC
生化的可以做pull down或者co-ip
【在 F*Q 的大作中提到】
: 外行向各位高手请教，多谢！
接触和作用。
比如一个vesicle上可以有很多蛋白，但他们不一定要直接接触。
要直接证明，还是需要其他手段，live imaging可以用， FRET或者BIFC
生化的可以做pull down或者co-ip
【在 F*Q 的大作中提到】
: 外行向各位高手请教，多谢！
F*Q
5 楼
很好，多谢了！
【在 b******s 的大作中提到】
: 很多co-localization只是同是定位于胞内的compartments，并不能说明两个蛋白相互
: 接触和作用。
: 比如一个vesicle上可以有很多蛋白，但他们不一定要直接接触。
: 要直接证明，还是需要其他手段，live imaging可以用， FRET或者BIFC
: 生化的可以做pull down或者co-ip
【在 b******s 的大作中提到】
: 很多co-localization只是同是定位于胞内的compartments，并不能说明两个蛋白相互
: 接触和作用。
: 比如一个vesicle上可以有很多蛋白，但他们不一定要直接接触。
: 要直接证明，还是需要其他手段，live imaging可以用， FRET或者BIFC
: 生化的可以做pull down或者co-ip
n*w
6 楼
most optical microscopes have a resolution >=200nm。
如果你要看的东西小于这个，有可能就只是空间上的交叉，不是真正意义上的
colocalization。。。
如果你要看的东西小于这个，有可能就只是空间上的交叉，不是真正意义上的
colocalization。。。
l*1
7 楼
he/she can try stimulated-emission depletion (STED)
paper:
htp://www.ncbi.nlm.nih.gov/pubmed/16896340
>We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy
with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is
demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell
Brand:
Super-Resolution Microscope Leica TCS STED
htp://www.leica-microsystems.com/products/confocal-microscopes/details/
product/leica-tcs-sted/
【在 n***w 的大作中提到】
: most optical microscopes have a resolution >=200nm。
: 如果你要看的东西小于这个，有可能就只是空间上的交叉，不是真正意义上的
: colocalization。。。
paper:
htp://www.ncbi.nlm.nih.gov/pubmed/16896340
>We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy
with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is
demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell
Brand:
Super-Resolution Microscope Leica TCS STED
htp://www.leica-microsystems.com/products/confocal-microscopes/details/
product/leica-tcs-sted/
【在 n***w 的大作中提到】
: most optical microscopes have a resolution >=200nm。
: 如果你要看的东西小于这个，有可能就只是空间上的交叉，不是真正意义上的
: colocalization。。。
g*5
8 楼
mark
s*j
9 楼
1.5M 一台. 美国也没几台吧?
emission depletion (STED) microscopy
attained in this study is
(ER) of a mammalian cell
【在 l**********1 的大作中提到】
: he/she can try stimulated-emission depletion (STED)
: paper:
: htp://www.ncbi.nlm.nih.gov/pubmed/16896340
: >We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy
: with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is
: demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell
: Brand:
: Super-Resolution Microscope Leica TCS STED
: htp://www.leica-microsystems.com/products/confocal-microscopes/details/
: product/leica-tcs-sted/
emission depletion (STED) microscopy
attained in this study is
(ER) of a mammalian cell
【在 l**********1 的大作中提到】
: he/she can try stimulated-emission depletion (STED)
: paper:
: htp://www.ncbi.nlm.nih.gov/pubmed/16896340
: >We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy
: with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is
: demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell
: Brand:
: Super-Resolution Microscope Leica TCS STED
: htp://www.leica-microsystems.com/products/confocal-microscopes/details/
: product/leica-tcs-sted/
F*Q
10 楼
70nm分辨率对于检测是否有contact还是不行吧？
emission depletion (STED) microscopy
attained in this study is
(ER) of a mammalian cell
【在 l**********1 的大作中提到】
: he/she can try stimulated-emission depletion (STED)
: paper:
: htp://www.ncbi.nlm.nih.gov/pubmed/16896340
: >We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy
: with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is
: demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell
: Brand:
: Super-Resolution Microscope Leica TCS STED
: htp://www.leica-microsystems.com/products/confocal-microscopes/details/
: product/leica-tcs-sted/
emission depletion (STED) microscopy
attained in this study is
(ER) of a mammalian cell
【在 l**********1 的大作中提到】
: he/she can try stimulated-emission depletion (STED)
: paper:
: htp://www.ncbi.nlm.nih.gov/pubmed/16896340
: >We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy
: with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is
: demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell
: Brand:
: Super-Resolution Microscope Leica TCS STED
: htp://www.leica-microsystems.com/products/confocal-microscopes/details/
: product/leica-tcs-sted/
l*1
11 楼
Pls go to
httq://www.ncbi.nlm.nih.gov/pubmed/21926998
NB: http now is "httq" from some bug might happened while using direct URL
address here.
if FAQ (Just F A Q) 的 target scale is below 15nm i.e. virus likes HSV
now your only choice is SEM not fluorescence microscopy.
pls refer
2011 paper:
httq://www.ncbi.nlm.nih.gov/pubmed/21345968
Ps:
这里也是现炒现卖 没有楼上的提示
还得不到
科普 偶本人 和whom want to know sth about size scale:
30 nm Small virus (Picornaviruses)
httq://en.wikibooks.org/wiki/Cell_Biology/Introduction/Cell_size
so here thanks so much to FAQ (Just F A Q) 的大作.
【在 F*Q 的大作中提到】
: 70nm分辨率对于检测是否有contact还是不行吧？
:
: emission depletion (STED) microscopy
: attained in this study is
: (ER) of a mammalian cell
httq://www.ncbi.nlm.nih.gov/pubmed/21926998
NB: http now is "httq" from some bug might happened while using direct URL
address here.
if FAQ (Just F A Q) 的 target scale is below 15nm i.e. virus likes HSV
now your only choice is SEM not fluorescence microscopy.
pls refer
2011 paper:
httq://www.ncbi.nlm.nih.gov/pubmed/21345968
Ps:
这里也是现炒现卖 没有楼上的提示
还得不到
科普 偶本人 和whom want to know sth about size scale:
30 nm Small virus (Picornaviruses)
httq://en.wikibooks.org/wiki/Cell_Biology/Introduction/Cell_size
so here thanks so much to FAQ (Just F A Q) 的大作.
【在 F*Q 的大作中提到】
: 70nm分辨率对于检测是否有contact还是不行吧？
:
: emission depletion (STED) microscopy
: attained in this study is
: (ER) of a mammalian cell
l*1
12 楼
就算最大的同步加速器全美就斯坦福那里有一台
需要做结构生物学实验的 能不从纽约飞加州去而做下来吗?
ZZ
比如 吴健雄
1956年，李政道、杨振宁在《物理评论》上发表了一篇著名的评论，他们在文章中提出
，有一项定律可能不成立。为了验证这一说法，他们与吴健雄商量，希望能用实验来给
予证实。
吴健雄在这方面称得上是权威，她建议用钴－60的衰变做实验。
当时正是严冬时节，实验场所设立在华盛顿市的国家标准局。为了完成试验，吴健
雄经常冒着风雪往返于纽约与华盛顿之间。功夫不负苦心人，试验结果终于出来了，这
一定律果然不成立。这一神奇的发现使李、杨获得了诺贝尔物理学奖
httq://www.shigu.com/modules/article/reader.php?aid=92&cid=2363
----
还有
httq://fds.duke.edu/db/aas/Physics/gao
在美国的学习研究生活中，高老师的导师也对她一直非常支持。有一段时间高老师在美国东部的MIT做实验，孩子和家
人都在西海岸的加州，高老师需要经常在相隔数千里的家庭和实验室之间。Bob知道此事后，赞助了每次往返的机票。
为此高老师对他非常感激。这种感激之情在高老师自己走上教职岗位，开始有自己的科研小组，并独立管理研究经费
后，更是有过之而无不及。几张机票这件事看似很小，但是在实验室各种开销中抽出这样一笔钱来帮助学生并不是那么
一件容易的事。
httq://daf.tsinghua.edu.cn/html/xinwen/jdxw/20101121/20385.html
【在 s*****j 的大作中提到】
: 1.5M 一台. 美国也没几台吧?
:
: emission depletion (STED) microscopy
: attained in this study is
: (ER) of a mammalian cell
需要做结构生物学实验的 能不从纽约飞加州去而做下来吗?
ZZ
比如 吴健雄
1956年，李政道、杨振宁在《物理评论》上发表了一篇著名的评论，他们在文章中提出
，有一项定律可能不成立。为了验证这一说法，他们与吴健雄商量，希望能用实验来给
予证实。
吴健雄在这方面称得上是权威，她建议用钴－60的衰变做实验。
当时正是严冬时节，实验场所设立在华盛顿市的国家标准局。为了完成试验，吴健
雄经常冒着风雪往返于纽约与华盛顿之间。功夫不负苦心人，试验结果终于出来了，这
一定律果然不成立。这一神奇的发现使李、杨获得了诺贝尔物理学奖
httq://www.shigu.com/modules/article/reader.php?aid=92&cid=2363
----
还有
httq://fds.duke.edu/db/aas/Physics/gao
在美国的学习研究生活中，高老师的导师也对她一直非常支持。有一段时间高老师在美国东部的MIT做实验，孩子和家
人都在西海岸的加州，高老师需要经常在相隔数千里的家庭和实验室之间。Bob知道此事后，赞助了每次往返的机票。
为此高老师对他非常感激。这种感激之情在高老师自己走上教职岗位，开始有自己的科研小组，并独立管理研究经费
后，更是有过之而无不及。几张机票这件事看似很小，但是在实验室各种开销中抽出这样一笔钱来帮助学生并不是那么
一件容易的事。
httq://daf.tsinghua.edu.cn/html/xinwen/jdxw/20101121/20385.html
【在 s*****j 的大作中提到】
: 1.5M 一台. 美国也没几台吧?
:
: emission depletion (STED) microscopy
: attained in this study is
: (ER) of a mammalian cell
l*1
13 楼
without your this 0F ask i never can catch above link URL address.
and some of them can be referenced on my next proposals.
so thanks so much to FAQ.
【在 F*Q 的大作中提到】
: 很好，多谢了！
and some of them can be referenced on my next proposals.
so thanks so much to FAQ.
【在 F*Q 的大作中提到】
: 很好，多谢了！
s*j
14 楼
STMD 是什么新东西?
practicalaspects.html
【在 l**********1 的大作中提到】
: without your this 0F ask i never can catch above link URL address.
: and some of them can be referenced on my next proposals.
: so thanks so much to FAQ.
practicalaspects.html
【在 l**********1 的大作中提到】
: without your this 0F ask i never can catch above link URL address.
: and some of them can be referenced on my next proposals.
: so thanks so much to FAQ.
l*1
15 楼
key board input miss
not M but E
STED
【在 s*****j 的大作中提到】
: STMD 是什么新东西?
:
: practicalaspects.html
not M but E
STED
【在 s*****j 的大作中提到】
: STMD 是什么新东西?
:
: practicalaspects.html
F*Q
16 楼
Sounds like we may find something to collaborate ...
【在 l**********1 的大作中提到】
: without your this 0F ask i never can catch above link URL address.
: and some of them can be referenced on my next proposals.
: so thanks so much to FAQ.
【在 l**********1 的大作中提到】
: without your this 0F ask i never can catch above link URL address.
: and some of them can be referenced on my next proposals.
: so thanks so much to FAQ.
l*1
17 楼
站内短信收到 已回复你了 不要人肉搜索这里阿
【在 F*Q 的大作中提到】
: Sounds like we may find something to collaborate ...
【在 F*Q 的大作中提到】
: Sounds like we may find something to collaborate ...
F*Q
18 楼
放心吧，那种宵小的行为，FAQ再闲也不会干的。
【在 l**********1 的大作中提到】
: 站内短信收到 已回复你了 不要人肉搜索这里阿
【在 l**********1 的大作中提到】
: 站内短信收到 已回复你了 不要人肉搜索这里阿
p*l
19 楼
FRET is the best way to confirm interaction, although FRET between more than 2 molecules can be very tricky.
Super resolution imaging techniques can give you at best 20~30 nm resolution
in biological samples. With super resolution imaging, the best you can get
is to narrow down two molecules within 20~30 nm range. That's assuming you have a perfect system, fine tuned and operated by an optical scientist. At 20~30nm resolution, saying two molecules are interacting is a very good guess, but not 100% bullet proof. To answer your question, you will also need multi-color super resolution imaging, which is much harder than single color. You'd better find a lab doing this to collaborate with if you want to go that way.
Like someone had pointed out, no matter what fluorescent imaging method you want to use, results from immunostain is not reliable. After two layers of antibody, the location of the fluorophore can be far away from the target.
Super resolution imaging techniques can give you at best 20~30 nm resolution
in biological samples. With super resolution imaging, the best you can get
is to narrow down two molecules within 20~30 nm range. That's assuming you have a perfect system, fine tuned and operated by an optical scientist. At 20~30nm resolution, saying two molecules are interacting is a very good guess, but not 100% bullet proof. To answer your question, you will also need multi-color super resolution imaging, which is much harder than single color. You'd better find a lab doing this to collaborate with if you want to go that way.
Like someone had pointed out, no matter what fluorescent imaging method you want to use, results from immunostain is not reliable. After two layers of antibody, the location of the fluorophore can be far away from the target.
p*l
20 楼
好的confocal也要80万，还是会有很多地方买得起的。
【在 s*****j 的大作中提到】
: STMD 是什么新东西?
:
: practicalaspects.html
【在 s*****j 的大作中提到】
: STMD 是什么新东西?
:
: practicalaspects.html
F*Q
21 楼
Two layers of antibody? Did you mean primary and secondary antibodies? Is it
general to have two layers of antibodies in fluorescent microscopy?
than 2 molecules can be very tricky.
resolution...
you want to use, results from immunostain is not reliable. After two layers
of antibody, the location of the fluorophore can be far away from the target
.
【在 p*l 的大作中提到】
: FRET is the best way to confirm interaction, although FRET between more than 2 molecules can be very tricky.
: Super resolution imaging techniques can give you at best 20~30 nm resolution
: in biological samples. With super resolution imaging, the best you can get
: is to narrow down two molecules within 20~30 nm range. That's assuming you have a perfect system, fine tuned and operated by an optical scientist. At 20~30nm resolution, saying two molecules are interacting is a very good guess, but not 100% bullet proof. To answer your question, you will also need multi-color super resolution imaging, which is much harder than single color. You'd better find a lab doing this to collaborate with if you want to go that way.
: Like someone had pointed out, no matter what fluorescent imaging method you want to use, results from immunostain is not reliable. After two layers of antibody, the location of the fluorophore can be far away from the target.
general to have two layers of antibodies in fluorescent microscopy?
than 2 molecules can be very tricky.
resolution...
you want to use, results from immunostain is not reliable. After two layers
of antibody, the location of the fluorophore can be far away from the target
.
【在 p*l 的大作中提到】
: FRET is the best way to confirm interaction, although FRET between more than 2 molecules can be very tricky.
: Super resolution imaging techniques can give you at best 20~30 nm resolution
: in biological samples. With super resolution imaging, the best you can get
: is to narrow down two molecules within 20~30 nm range. That's assuming you have a perfect system, fine tuned and operated by an optical scientist. At 20~30nm resolution, saying two molecules are interacting is a very good guess, but not 100% bullet proof. To answer your question, you will also need multi-color super resolution imaging, which is much harder than single color. You'd better find a lab doing this to collaborate with if you want to go that way.
: Like someone had pointed out, no matter what fluorescent imaging method you want to use, results from immunostain is not reliable. After two layers of antibody, the location of the fluorophore can be far away from the target.
p*l
22 楼
Indirect label is more common.
l*1
23 楼
the ultra small scale below 18nm
you should try
AFM Atomic Force microscopy
paper link:
//www.ncbi.nlm.nih.gov/pubmed/20202756
from below lab:
//www.ntnu.edu/employees/dionnecg
【在 F*Q 的大作中提到】
: 放心吧，那种宵小的行为，FAQ再闲也不会干的。
you should try
AFM Atomic Force microscopy
paper link:
//www.ncbi.nlm.nih.gov/pubmed/20202756
from below lab:
//www.ntnu.edu/employees/dionnecg
【在 F*Q 的大作中提到】
: 放心吧，那种宵小的行为，FAQ再闲也不会干的。
s*g
24 楼
bingo! FRET is the answer
STED也就是个bubble,仪器又贵又不适用,UCLA3年前就有了,但是现在也没派上大用处.
我看有前景的就是SIM和STORM/PALM,后者更有前景,如果没钱自己都可以搭起来一套.
than 2 molecules can be very tricky.
resolution
get
have a perfect system, fine tuned and operated by an optical scientist. At
20~30nm resolution, saying two molecules are interacting is a very good
guess, but not 100% bullet proof. To answer your question, you will also
need multi-color super resolution imaging, which is much harder than single
color. You'd better find a lab doing this to collaborate with if you want to
go that way.
you want to use, results from immunostain is not reliable. After two layers
of antibody, the location of the fluorophore can be far away from the target
.
【在 p*l 的大作中提到】
: FRET is the best way to confirm interaction, although FRET between more than 2 molecules can be very tricky.
: Super resolution imaging techniques can give you at best 20~30 nm resolution
: in biological samples. With super resolution imaging, the best you can get
: is to narrow down two molecules within 20~30 nm range. That's assuming you have a perfect system, fine tuned and operated by an optical scientist. At 20~30nm resolution, saying two molecules are interacting is a very good guess, but not 100% bullet proof. To answer your question, you will also need multi-color super resolution imaging, which is much harder than single color. You'd better find a lab doing this to collaborate with if you want to go that way.
: Like someone had pointed out, no matter what fluorescent imaging method you want to use, results from immunostain is not reliable. After two layers of antibody, the location of the fluorophore can be far away from the target.
STED也就是个bubble,仪器又贵又不适用,UCLA3年前就有了,但是现在也没派上大用处.
我看有前景的就是SIM和STORM/PALM,后者更有前景,如果没钱自己都可以搭起来一套.
than 2 molecules can be very tricky.
resolution
get
have a perfect system, fine tuned and operated by an optical scientist. At
20~30nm resolution, saying two molecules are interacting is a very good
guess, but not 100% bullet proof. To answer your question, you will also
need multi-color super resolution imaging, which is much harder than single
color. You'd better find a lab doing this to collaborate with if you want to
go that way.
you want to use, results from immunostain is not reliable. After two layers
of antibody, the location of the fluorophore can be far away from the target
.
【在 p*l 的大作中提到】
: FRET is the best way to confirm interaction, although FRET between more than 2 molecules can be very tricky.
: Super resolution imaging techniques can give you at best 20~30 nm resolution
: in biological samples. With super resolution imaging, the best you can get
: is to narrow down two molecules within 20~30 nm range. That's assuming you have a perfect system, fine tuned and operated by an optical scientist. At 20~30nm resolution, saying two molecules are interacting is a very good guess, but not 100% bullet proof. To answer your question, you will also need multi-color super resolution imaging, which is much harder than single color. You'd better find a lab doing this to collaborate with if you want to go that way.
: Like someone had pointed out, no matter what fluorescent imaging method you want to use, results from immunostain is not reliable. After two layers of antibody, the location of the fluorophore can be far away from the target.
l*1
25 楼
Really?
after you read below Nature Methods paper
its link:
on
http://www.mitbbs.com/article_t/Biology/31634159.html
then might had different view?
At
single
【在 s*****g 的大作中提到】
: bingo! FRET is the answer
: STED也就是个bubble,仪器又贵又不适用,UCLA3年前就有了,但是现在也没派上大用处.
: 我看有前景的就是SIM和STORM/PALM,后者更有前景,如果没钱自己都可以搭起来一套.
:
: than 2 molecules can be very tricky.
: resolution
: get
: have a perfect system, fine tuned and operated by an optical scientist. At
: 20~30nm resolution, saying two molecules are interacting is a very good
: guess, but not 100% bullet proof. To answer your question, you will also
after you read below Nature Methods paper
its link:
on
http://www.mitbbs.com/article_t/Biology/31634159.html
then might had different view?
At
single
【在 s*****g 的大作中提到】
: bingo! FRET is the answer
: STED也就是个bubble,仪器又贵又不适用,UCLA3年前就有了,但是现在也没派上大用处.
: 我看有前景的就是SIM和STORM/PALM,后者更有前景,如果没钱自己都可以搭起来一套.
:
: than 2 molecules can be very tricky.
: resolution
: get
: have a perfect system, fine tuned and operated by an optical scientist. At
: 20~30nm resolution, saying two molecules are interacting is a very good
: guess, but not 100% bullet proof. To answer your question, you will also
s*g
26 楼
还没读,先去读一下
【在 l**********1 的大作中提到】
: Really?
: after you read below Nature Methods paper
: its link:
: on
: http://www.mitbbs.com/article_t/Biology/31634159.html
: then might had different view?
:
: At
: single
【在 l**********1 的大作中提到】
: Really?
: after you read below Nature Methods paper
: its link:
: on
: http://www.mitbbs.com/article_t/Biology/31634159.html
: then might had different view?
:
: At
: single
F*Q
27 楼
Besides the resolution issue, is it guaranteed that no artifacts in
immunolabeling in live cell imaging?
【在 l**********1 的大作中提到】
: Really?
: after you read below Nature Methods paper
: its link:
: on
: http://www.mitbbs.com/article_t/Biology/31634159.html
: then might had different view?
:
: At
: single
immunolabeling in live cell imaging?
【在 l**********1 的大作中提到】
: Really?
: after you read below Nature Methods paper
: its link:
: on
: http://www.mitbbs.com/article_t/Biology/31634159.html
: then might had different view?
:
: At
: single
p*l
28 楼
STED is not a bubble. It has limitations, but PALM and STORM have too. The
strength of STED is live animal imaging not cell imaging. Hell has a
Science paper coming up about live mouse brain imaging.
For regular microscope-type application, SIM is more user friendly, but it
cannot reach beyond 100 nm on biological samples.
At
single
【在 s*****g 的大作中提到】
: bingo! FRET is the answer
: STED也就是个bubble,仪器又贵又不适用,UCLA3年前就有了,但是现在也没派上大用处.
: 我看有前景的就是SIM和STORM/PALM,后者更有前景,如果没钱自己都可以搭起来一套.
:
: than 2 molecules can be very tricky.
: resolution
: get
: have a perfect system, fine tuned and operated by an optical scientist. At
: 20~30nm resolution, saying two molecules are interacting is a very good
: guess, but not 100% bullet proof. To answer your question, you will also
strength of STED is live animal imaging not cell imaging. Hell has a
Science paper coming up about live mouse brain imaging.
For regular microscope-type application, SIM is more user friendly, but it
cannot reach beyond 100 nm on biological samples.
At
single
【在 s*****g 的大作中提到】
: bingo! FRET is the answer
: STED也就是个bubble,仪器又贵又不适用,UCLA3年前就有了,但是现在也没派上大用处.
: 我看有前景的就是SIM和STORM/PALM,后者更有前景,如果没钱自己都可以搭起来一套.
:
: than 2 molecules can be very tricky.
: resolution
: get
: have a perfect system, fine tuned and operated by an optical scientist. At
: 20~30nm resolution, saying two molecules are interacting is a very good
: guess, but not 100% bullet proof. To answer your question, you will also
h*9
29 楼
至少10A级别左右的吧。70nm都到火星去了。
【在 F*Q 的大作中提到】
: 70nm分辨率对于检测是否有contact还是不行吧？
:
: emission depletion (STED) microscopy
: attained in this study is
: (ER) of a mammalian cell
【在 F*Q 的大作中提到】
: 70nm分辨率对于检测是否有contact还是不行吧？
:
: emission depletion (STED) microscopy
: attained in this study is
: (ER) of a mammalian cell
F*Q
30 楼
这个还是比较tricky。有没有关于contact的确切定义？比如说是否需要有某种化学键
的存在来证明interaction？也许有contact的两个蛋白或结构也不一定就存在相互作用
（某种化学键的存在）吧？
是否需要in vitro的binding来验证是否有interaction?
【在 h*****9 的大作中提到】
: 至少10A级别左右的吧。70nm都到火星去了。
的存在来证明interaction？也许有contact的两个蛋白或结构也不一定就存在相互作用
（某种化学键的存在）吧？
是否需要in vitro的binding来验证是否有interaction?
【在 h*****9 的大作中提到】
: 至少10A级别左右的吧。70nm都到火星去了。
l*1
31 楼
需要质子或电子和异类的原子核的高能物理实验来验证是否有interaction吧? 极端的
话 上中微子实验验证啊--trigger
//zh.wikipedia.org/wiki/歐洲核子研究組織
【在 F*Q 的大作中提到】
: 这个还是比较tricky。有没有关于contact的确切定义？比如说是否需要有某种化学键
: 的存在来证明interaction？也许有contact的两个蛋白或结构也不一定就存在相互作用
: （某种化学键的存在）吧？
: 是否需要in vitro的binding来验证是否有interaction?
话 上中微子实验验证啊--trigger
//zh.wikipedia.org/wiki/歐洲核子研究組織
【在 F*Q 的大作中提到】
: 这个还是比较tricky。有没有关于contact的确切定义？比如说是否需要有某种化学键
: 的存在来证明interaction？也许有contact的两个蛋白或结构也不一定就存在相互作用
: （某种化学键的存在）吧？
: 是否需要in vitro的binding来验证是否有interaction?
F*Q
32 楼
80万要说多也多，要说不多也不多。当年的老板为了留我，花了差不多那么多钱订购了
一台仪器外加一些辅助设备，最后因为我的结果不支持他以前发表的一些文章而明确表
达他不能发表我的结果，我只好走人。在我走的时候定购的仪器还没到货，仪器到了之
后因为实验室没有人做那个方向了，而且他也不打算自己恶心自己再去碰那个方向，只
好把从没用过的新仪器拱手送给别人了。悲哀吧？
【在 p*l 的大作中提到】
: 好的confocal也要80万，还是会有很多地方买得起的。
一台仪器外加一些辅助设备，最后因为我的结果不支持他以前发表的一些文章而明确表
达他不能发表我的结果，我只好走人。在我走的时候定购的仪器还没到货，仪器到了之
后因为实验室没有人做那个方向了，而且他也不打算自己恶心自己再去碰那个方向，只
好把从没用过的新仪器拱手送给别人了。悲哀吧？
【在 p*l 的大作中提到】
: 好的confocal也要80万，还是会有很多地方买得起的。
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