Embryos imaging

[Vladimir Ghukasyan-2]

Dear List members,

We're trying to do blastocysts confocal imaging but face a problem of
active movement. They are pretty reluctant to adhesions, hence the
difficulty with immobilization. Did you face the same problem, and if
yes - what would be the suggestion to try to overcome it?

Thank you in advance,
Vladimir

[Alice Rodriguez Diaz]

Dear Vladimir,

Fast imaging with a spinning disk confocal. And Teflon film to allow air and
movement.. ?

Alice

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Vladimir Gukassyan
Sent: Friday, May 21, 2010 1:14 PM
To: [hidden email]
Subject: Embryos imaging

Dear List members,

We're trying to do blastocysts confocal imaging but face a problem of
active movement. They are pretty reluctant to adhesions, hence the
difficulty with immobilization. Did you face the same problem, and if
yes - what would be the suggestion to try to overcome it?

Thank you in advance,
Vladimir

[Vladimir Ghukasyan-2]

Dear Alice,

Thank you for your reply.
Unfortunately, we have only LSM confocal. Could you clarify about the
Teflon film please?

With regards,
Vladimir


On Fri, May 21, 2010 at 2:29 PM, Alice Rodriguez Diaz
<[hidden email]> wrote:

> Dear Vladimir,
>
> Fast imaging with a spinning disk confocal. And Teflon film to allow air and
> movement.. ?
>
> Alice
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On
> Behalf Of Vladimir Gukassyan
> Sent: Friday, May 21, 2010 1:14 PM
> To: [hidden email]
> Subject: Embryos imaging
>
> Dear List members,
>
> We're trying to do blastocysts confocal imaging but face a problem of
> active movement. They are pretty reluctant to adhesions, hence the
> difficulty with immobilization. Did you face the same problem, and if
> yes - what would be the suggestion to try to overcome it?
>
> Thank you in advance,
> Vladimir
>

[Stephen Cody-2]

Dear Vladimir,

Try mounting in low melting point agarose. To dissolve in your buffer  
you need to heat to about 60 deg C. But then you can cool it down to  
37deg C and it will stay molten until cooled to 35 deg C. Once  
solidified you can warm to 37deg again. It is perfect for imaging  
embryos.

Steve

Stephen H. Cody

On 22/05/2010, at 5:05 AM, Vladimir Gukassyan <[hidden email]>  
wrote:

> Dear Alice,
>
> Thank you for your reply.
> Unfortunately, we have only LSM confocal. Could you clarify about the
> Teflon film please?
>
> With regards,
> Vladimir
>
>
> On Fri, May 21, 2010 at 2:29 PM, Alice Rodriguez Diaz
> <[hidden email]> wrote:
>> Dear Vladimir,
>>
>> Fast imaging with a spinning disk confocal. And Teflon film to  
>> allow air and
>> movement.. ?
>>
>> Alice
>>
>> -----Original Message-----
>> From: Confocal Microscopy List  
>> [mailto:[hidden email]] On
>> Behalf Of Vladimir Gukassyan
>> Sent: Friday, May 21, 2010 1:14 PM
>> To: [hidden email]
>> Subject: Embryos imaging
>>
>> Dear List members,
>>
>> We're trying to do blastocysts confocal imaging but face a problem of
>> active movement. They are pretty reluctant to adhesions, hence the
>> difficulty with immobilization. Did you face the same problem, and if
>> yes - what would be the suggestion to try to overcome it?
>>
>> Thank you in advance,
>> Vladimir
>>

[Barry O'Brien]

Infiltration with glycerol over a couple of days helps.  Temperature
equilibration is also an area to pay attention to.

If anyone has a reliable method of stabilizing the embryo after
orienting it while on the microscope stage then we would also like to
hear about it.

Thanks,

Barry

At 06:13 a.m. 22/05/2010, you wrote:
>Dear List members,
>
>We're trying to do blastocysts confocal imaging but face a problem of
>active movement. They are pretty reluctant to adhesions, hence the
>difficulty with immobilization. Did you face the same problem, and if
>yes - what would be the suggestion to try to overcome it?
>
>Thank you in advance,
>Vladimir

Dr Barry O'Brien
Dept of Biological Sciences,
University of Waikato
Private Bag 3105
HAMILTON
New Zealand

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