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Question to be answered please

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Question to be answered please

Postby hashemyemen » Sat Mar 20, 2010 5:48 pm

I would like to know what type of microscope should be used during performing " animals' cloning ", I mean in that step of nucleus isolation and injection !!

Can i use Light microscope or I must use other type than that??


Regards.....
Last edited by hashemyemen on Sat Mar 20, 2010 6:10 pm, edited 1 time in total.
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Postby JackBean » Sat Mar 20, 2010 5:52 pm

You, evolution blamer, wants to clone animals?

Some fine light microscope should be fine

I thought, that you have already cloned animals... :roll: :-O
post-119123.html#p119123
http://www.biolib.cz/en/main/

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Re:

Postby hashemyemen » Sat Mar 20, 2010 6:06 pm

JackBean wrote:You, evolution blamer, wants to clone animals?

Some fine light microscope should be fine

I thought, that you have already cloned animals... :roll: :-O
post-119123.html#p119123


No, actually I usually use Electron Microscope due to its high magnification, but I want to try this time via light microscope....

By the way, How can you say " I thought, that you have already cloned animals "..
Sweetie, my previous project about blocking transofmration of genetic information was via PCR technology, and it was not at all dealing with cloning. Means i was dealing with molecular biology not cell biology...

one thing els, I hope that you guide me to the best way of microinjection as well.... :D

and thank you very much
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Postby JackBean » Sat Mar 20, 2010 7:59 pm

Well, IMHO if you changed their DNA, you had to transform their cells, didn't you?
http://www.biolib.cz/en/main/

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Postby david23 » Sat Mar 20, 2010 10:11 pm

If you are using it for the injection then it's live sample, something that EM cant do.
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Re:

Postby hashemyemen » Sun Mar 21, 2010 1:58 am

JackBean wrote:Well, IMHO if you changed their DNA, you had to transform their cells, didn't you?


Yes, Of coure.
But that procedure is simple, because I did changed their DNA via rDNA technology, then after the induction of cellular division, I tranformed the changed DNA again to the animal......
That experiment was something other than what I'm about to do now. By that way, L want to know what are the suitable conditions for maintains Somatic cell under the terms of Lab ??
I mean who much Ph, and temperature?
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Re:

Postby hashemyemen » Sun Mar 21, 2010 2:00 am

david23 wrote:If you are using it for the injection then it's live sample, something that EM cant do.


I think E.M can do it, because the cells are placed in the culture medium while transforming the nucleus from cell to another cell...
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Re: Re:

Postby biohazard » Mon Mar 22, 2010 8:01 am

hashemyemen wrote:
david23 wrote:If you are using it for the injection then it's live sample, something that EM cant do.


I think E.M can do it, because the cells are placed in the culture medium while transforming the nucleus from cell to another cell...


You cannot use EM in microinjection for several reasons: it kills cells, which is a pretty obvious reason. Depending on whether you use SEM or TEM or something else, you must fix and then coat or stain the cells/target organisms in order to get the image. Furthermore the cells are typically placed in a high vacuum, something which live cells do not like. You also cannot manipulate the cells, that is, perform the injection under the EM. Lastly, you do not even need an EM for this because a light microscope is sufficient to visualise animal cells, just like JackBean already said.
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Re: Re:

Postby hashemyemen » Mon Mar 22, 2010 11:47 am

biohazard wrote:
hashemyemen wrote:
david23 wrote:If you are using it for the injection then it's live sample, something that EM cant do.


I think E.M can do it, because the cells are placed in the culture medium while transforming the nucleus from cell to another cell...


You cannot use EM in microinjection for several reasons: it kills cells, which is a pretty obvious reason. Depending on whether you use SEM or TEM or something else, you must fix and then coat or stain the cells/target organisms in order to get the image. Furthermore the cells are typically placed in a high vacuum, something which live cells do not like. You also cannot manipulate the cells, that is, perform the injection under the EM. Lastly, you do not even need an EM for this because a light microscope is sufficient to visualise animal cells, just like JackBean already said.


Hmmmmmm, You are right up to some extent because most of my trying in this protocol was so much success, yet I injected the nucleus and transformed it, but no good results !!

Hmmm, I hope that you give full idea about utilizing L.Microscope for micro-injection procedure...


Image

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My question is:

Do we need this high developed microscope :

Image

Or we can use this simple as well??

Image

:?
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Postby JackBean » Mon Mar 22, 2010 12:21 pm

IMHO in the simple one you
1) don't have enough space
2) your hand is not from stone, so it would be probably better, if the injection was hold by machine...
http://www.biolib.cz/en/main/

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Postby biohazard » Mon Mar 22, 2010 12:25 pm

It is mostly about the technical skill of the operator. A skilled person with good injection tools might be able to carry out the procedure even under a fairly basic microscope. Actually the only required thing from the microscope is that it allows at least around x400 magnification and has enough light output to give clear vision and there is enough space to operate the petri dish and the injector on the sample holding space. Actually much bigger magnification is not even practically possible, since immersion oil would be required. A skilled person can perform the injection even with smaller magnifications than x400.
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Re: Re:

Postby JackBean » Mon Mar 22, 2010 12:29 pm

hashemyemen wrote:But that procedure is simple, because I did changed their DNA via rDNA technology, then after the induction of cellular division, I tranformed the changed DNA again to the animal......

that's my point, you had to transform your cells, so how did you do that, as you had transfered the change to offspring too
http://www.biolib.cz/en/main/

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