cryoprotectant

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Arvind Singh Pundir
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cryoprotectant

how long can one keep the tissue in 30% sucrose with sodium azide without any damage to the tissue if there is delay in cryosectioning

heehawmcduff
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I've known people in my

I've known people in my previous lab to store up to 3 days but I understand that it may be possible to go longer.

Arvind Singh Pundir
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heehawmcduff wrote:

heehawmcduff wrote:

I've known people in my previous lab to store up to 3 days but I understand that it may be possible to go longer.

thanks
its the time tissue takes to sink in sucrose, can you find out with your previous contacts how long can the tissue be stored without damage to the tissue

Guest (not verified)
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1) I have stored blocks for

1) I have stored blocks for up to 1 year, it depends on your antigen.
2) it is very common with large brains (non-human primate) to leave them until they sink. This may take a week.

heehawmcduff
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Pundir - I'll get in touch

Pundir - I'll get in touch with people in my old lab and see what info they have.
Best wishes

Arvind Singh Pundir
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heehawmcduff wrote:

heehawmcduff wrote:

Pundir - I'll get in touch with people in my old lab and see what info they have.
Best wishes

 Dear  heehawmcduff 
 
please also ask them what they are using as preservative or agent to avoid any growth as i am using sodium azide 0.01% in my case
thanks
 

heehawmcduff
heehawmcduff's picture
Hi

Hi
So apparently a chap in my old lab used to use sucrose to inflate lungs and then freeze them before they were sectioned.  At -20, with sodium azide added too, he kept them for up to 6 months.  He said that if you did keep lung tissue for too long then it would become unusable (because of the delicate architecture) so I guess it very much depends on the tissue. 
I'm sorry I can't be more helpful - not much experience with cryosectioning.

Jason King
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I used to do cryosectioning

I used to do cryosectioning of mouse lungs and inflated the lungs with an OCT/sucrose mixture then froze in liquid nitrogen. Stored at -80C. I did not use an antimicrobial such as Na azide as I was storing samples at -80C. I never tested how long it was possible to leave the lungs in this state of preservation but I wouldn't expect much to change during storage. It is far more critical to get the cryosectioning conditions right; eg. cabinet temperature, blade temperature, sharp blade etc.

heehawmcduff
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Probably a silly followup

Probably a silly followup question, but did you see any distortion of the murine lung tissue when storing at such low temps or did the sucrose/oct work well to keep them intact?
Regards

Arvind Singh Pundir
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heehawmcduff wrote:

heehawmcduff wrote:

Probably a silly followup question, but did you see any distortion of the murine lung tissue when storing at such low temps or did the sucrose/oct work well to keep them intact?
Regards

 Hi heehawmcduff
prabably theOCT provide it a appropriate texture to cut the tissue without damage to the tissue rest is how you manipulate the temp  for sectioning as a lot depends on temp at which you section the tissue
 

heehawmcduff
heehawmcduff's picture
Thanks very much for that

Thanks very much for that reply - it's something I'd like to do a little more so will no doubt be back with a few other queries :)