Help for spleen frozen section

[Lucyusf]

Hi, everybody,

I'm new to spleen section. I tried same thing as brain sectioning. 4% paraformadehyde 24h, 10% sucrose 24h, 20% sucrose 24h and 30% sucrose 24h, then cutting at -20 degree. Unfortunately, I got a lot of brown thing on the slide even with hematoxylin staining. I tried to attach my picture, but it didn't work.

Can anybody help me with the problem?

Thank You very much.

Lucy

[excalibur]

Spleen is bloody and shatters easily, even in paraffin. Try cutting warmer or holding your thumb on the block face a sec and then quickly cut.

[Lucyusf]

Thanks a lot. I'll try to increase the temperature.

Lucy

[Lucyusf]

Attached is the spleen stained with hematoxylin alone.

[Yao]

Attached is the spleen stained with hematoxylin alone.

are those brown things erythrocytes, can they be stained by eosin

[gula]

I guess this is hemosiderin from the destroyed old erythrocytes in the spleen. This is a common and non-pathalogic pigment after decomposition of hemoglobin.
Try a prussian-blue stain for demonstration of iron.
Compare these pictures with your section:
http://www.pathology.vcu.edu/education/gi/lab3.c.html

bye gula

[luza123]

Sorry,I can't help you.