Cryosection

7 posts / 0 new
Last post
Jack and Jill
Jack and Jill's picture
Cryosection

Hi everyone. I am facing problem with immunostaining on my cryo-sectioning slides. I have checked all the steps in my present staining protocol. It looks like there must be something went wrong between after sectioning and before staining. Here is what I have done. After collecting the sections on my slides, I let them dry in the room temperature for 30min and stored them at -20 for 2 weeks. Before staining, I let them dry in the room tempereature for an hour. Is there anything wrong in the way I have done? I would appreciate any suggestion that could improve the situation
Thanks in advance

Guy Sovak
Guy Sovak's picture
Hi there and welcome to

Hi there and welcome to scientist solutions.
I would be more then happy to help you find a solution.
But, I would need to know more details. What sample are you looking to stain. What kind of Ab's you are using etc. Please let me know all the steps.
Guy

Marina Fomin
Marina Fomin's picture
Do you fix tissue before

Do you fix tissue before freezing or do you fix slides before staining?

Guy Sovak
Guy Sovak's picture
Marina Fomin wrote:Do you fix

Marina Fomin wrote:

Do you fix tissue before freezing or do you fix slides before staining?

The fixing should be done after freezing.
In Some exceptions you can do it before.
Guy

Marina Fomin
Marina Fomin's picture
guy wrote:Marina Fomin wrote

guy wrote:

Marina Fomin wrote:
Do you fix tissue before freezing or do you fix slides before staining?

The fixing should be done after freezing.
In Some exceptions you can do it before.
Guy

Actually, I fix tissue before freezing and it worked for most of samples.

Guy Sovak
Guy Sovak's picture
There are two main options to

There are two main options to to treat a sample before freezing it,
1) cut it put it in NBF / EDTA (for bone) and Freez 2) cut and freeze.
Usually I took the sample in my case bone put it in EDTA for 72 hours and freezed it.
Guy

Arvind Singh Pundir
Arvind Singh Pundir's picture
wrote:

wrote:

Hi you can try antigen retrieval as there is no prob with the procedure you are applying