Hi experts,
I have been reading a lot of articles from Abcam, Abnova, Thermo Fisher, etc. about the protocol for IHC. The method seems pretty simple and straightforward. But as I go along the way, I noticed a lot of bumps and hurdles along the way. I am working with frozen brain tissue sections.
Could you share your experience in cryosectioning, IHC, microscopy by answering these series of questions:
1. What is the method? (i.e. Cryosection, IHC, microscopy, etc.)
2. What organism? (i.e. mouse, pig, human, primate, etc.)
3. What is your sample? (i.e. brain, heart, bone marrow, etc.)
4. What is your target? (i.e. gene, protein, etc.) What kind of section? (i.e. coronal, sagittal, transverse) If applicable, what is the method of preservation? (i.e. instant freeze, paraffin embedded, etc.)
5. What was your experience or problem encountered? Provide details.
6. Did you solve it? How did you solve it? (Details)
By sharing your experience, researchers who have trouble or problems with this method can be easily solved. Thanks in advance!
1. IHC
2. mouse
3. brain
4.coronal, frozen tissue
5. During the first washing step, after tissue section/slices attached to adhesion slides, some tissues detach from the slides. 3 slices per slide. (Protocol here)
6. Not solved yet.
http://www.abcam.com/protocols/immunostaining-paraffin-frozen-free-floating-protocol
Thanks, @Yusmaris for your help. I realized that too. I am new about this cryosectioning, so your information is helpful. We have two types of adhesion slides I will post the photos later.
One slide has the (+) label at the edge of the slide (Fisher Scientific), one has some different labeling at the corner (Thermo).
I mistakenly used the latter, which I realized must be coated with the poly-L-lysine. Although some tissues attached, most of them come off.
One of our lab members pointed out that the first type of slide (with a + label at the corner) doesn't need a poly-L-lysine because it is charged and it will hold the tissues better. What do you think about this?
Thanks again. I will do that. But for now, seems like the (+)-charged slides are better than these slides (charged vs. non-charged; left vs. right; top bottom). I did them side by side with same tissue section. And the (+) charged slides hold them better than the other one (without poly-L-lysine). I appreciate your suggestion; I will try it later.
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