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Help-cell fixation problem
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Help-cell fixation problem# Biology - 生物学
F*w
1
父亲今年55岁,年纪不大,不过因为身体原因最近几年一直没有工作,连退休证也没有
。想问问大家这样的情况,如果签证官问起,要怎么回答?
还有一个问题是,这两天有人预约签证吗?每次我一拨通到选择语言那里就断了。。也
有人遇到类似的问题吗?
谢谢!!
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h*y
2
看过死人经,现在正看拔魔。两部开头都有些类似,小孩子们为了各种利益互相划小圈
子刻画的入木三分。大家小时候都有过类似的经历吧?
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y*6
3
HEK293 cells in 96-well plate, DMEM with 10% FBS, cultured 2d, about 100%
confluency.
Washed once with PBS before adding fixation solution (3.7% formaldehyde/PBS).
Cells detached in fixastion solution. Is it because formaldehyde does not
work with HEK293 cells? Anyone tried methenol? What's the concentration?
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h*e
4
Not sure about the answer to your first question.
But regards to making appointment, I called to make an interview a couple of
days ago. The phone system seemed fine to me. No problem.
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j*u
5
有人的地方就有江湖……

【在 h******y 的大作中提到】
: 看过死人经,现在正看拔魔。两部开头都有些类似,小孩子们为了各种利益互相划小圈
: 子刻画的入木三分。大家小时候都有过类似的经历吧?

avatar
h*y
6
Do not wash your cells with PBS as 293 cells are easy to detach. Add
fixative after remove medium, or pre-coat your plate with poly-D/L-Lysine.

PBS).

【在 y****6 的大作中提到】
: HEK293 cells in 96-well plate, DMEM with 10% FBS, cultured 2d, about 100%
: confluency.
: Washed once with PBS before adding fixation solution (3.7% formaldehyde/PBS).
: Cells detached in fixastion solution. Is it because formaldehyde does not
: work with HEK293 cells? Anyone tried methenol? What's the concentration?

avatar
F*w
7
谢谢楼上的回答!!
有人知道第一个问题要怎么说嘛?
谢谢了。
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f*0
8
等到300多章,你会发现,还是那群孩子在说话,没有成长。
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V*n
9
ice cold acetone
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h*y
10
目前手机上只看到170多章,剩下的等全本了再看

【在 f*****0 的大作中提到】
: 等到300多章,你会发现,还是那群孩子在说话,没有成长。
avatar
y*6
11
Can anyone explain how fixation works? My understanding is that fixative
helps the crosslink of proteins and other macromolecules. I also have an
impression that it helps the cells to attach to the plate more firmly. It is
correct?
avatar
b*e
12
拔魔我觉得挺失败的,起码我没有任何跟的欲望,虽然积攒多了有空会拿来消磨。
冰神最好的还是死人经前半部。拔魔和死人经后半部差不多,聊不起太多阅读欲望。

【在 h******y 的大作中提到】
: 目前手机上只看到170多章,剩下的等全本了再看
avatar
c*n
13
I use 4% Paraformaldehyde to fix 293 HEK cells for 15 min.
I always put cells on cover slips coated with fibronectin or polyL lysin and
then fix them. It works fine.

PBS).

【在 y****6 的大作中提到】
: HEK293 cells in 96-well plate, DMEM with 10% FBS, cultured 2d, about 100%
: confluency.
: Washed once with PBS before adding fixation solution (3.7% formaldehyde/PBS).
: Cells detached in fixastion solution. Is it because formaldehyde does not
: work with HEK293 cells? Anyone tried methenol? What's the concentration?

avatar
x*8
14
死人经基本以报仇为主线,比较单一,
拔魔看起来像是一部构思宏大的作品,似乎要阐述魔,道,人,妖的关系之类的,有一
个故事情节之上的主题要表述,现在可能刚刚展开,

【在 b*********e 的大作中提到】
: 拔魔我觉得挺失败的,起码我没有任何跟的欲望,虽然积攒多了有空会拿来消磨。
: 冰神最好的还是死人经前半部。拔魔和死人经后半部差不多,聊不起太多阅读欲望。

avatar
h*n
15
Fixation reagent is no use to help cells attch on a plate. I suggest coating
your
plate with poly-lysine

is

【在 y****6 的大作中提到】
: Can anyone explain how fixation works? My understanding is that fixative
: helps the crosslink of proteins and other macromolecules. I also have an
: impression that it helps the cells to attach to the plate more firmly. It is
: correct?

avatar
j*u
16
nod

【在 x**8 的大作中提到】
: 死人经基本以报仇为主线,比较单一,
: 拔魔看起来像是一部构思宏大的作品,似乎要阐述魔,道,人,妖的关系之类的,有一
: 个故事情节之上的主题要表述,现在可能刚刚展开,

avatar
b*l
17
不要用 PBS,因为里面没 Ca/Mg,会导致细胞 detach。
用 DPBS with Ca/Mg 来洗细胞,然后再 fix。
而且 PBS 太 hush。原则是活细胞用 DPBS,死细胞或者不关细胞才用 PBS。

PBS).

【在 y****6 的大作中提到】
: HEK293 cells in 96-well plate, DMEM with 10% FBS, cultured 2d, about 100%
: confluency.
: Washed once with PBS before adding fixation solution (3.7% formaldehyde/PBS).
: Cells detached in fixastion solution. Is it because formaldehyde does not
: work with HEK293 cells? Anyone tried methenol? What's the concentration?

avatar
z*4
18
+1
总结的太对了,死人最后看不下去了,同样故事反复说

【在 f*****0 的大作中提到】
: 等到300多章,你会发现,还是那群孩子在说话,没有成长。
avatar
y*6
19
Thanks a lot for the reply from all of you. I will try coating.
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