Cryosection [View Printable]
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Jack and Jill
Group: Member
Posts: 1
Joined: Aug 08, 2008
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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 |
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| Posted Aug 08, 2008, 18:30 PM |
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guy
Group: Admin
Posts: 340
Joined: Nov 28, 2005
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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 |
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| Posted Aug 09, 2008, 21:59 PM |
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Marina Fomin
Group: Member
Posts: 46
Joined: Feb 28, 2006
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Do you fix tissue before freezing or do you fix slides before staining? |
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| Posted Aug 19, 2008, 17:19 PM |
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guy
Group: Admin
Posts: 340
Joined: Nov 28, 2005
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| Marina Fomin said: | | 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 |
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| Posted Aug 20, 2008, 15:29 PM |
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Marina Fomin
Group: Member
Posts: 46
Joined: Feb 28, 2006
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| guy said: | | Marina Fomin said: | | 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. |
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| Posted Aug 20, 2008, 15:45 PM |
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guy
Group: Admin
Posts: 340
Joined: Nov 28, 2005
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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 |
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| Posted Aug 21, 2008, 11:01 AM |
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