Hi everyone!
I made a Perl's stain for some samples in my lab; they are old samples and I´m not sure how many time they spended on fixative (formol) or whatsoever, so I´m starting from the specimen embedded in paraffin.
5 microns thick sections of both liver and spleen, we wanted to show the hemosiderin on "germinal centers" on the spleen (that it shows) but we also expected some positives on the liver due to erythrocyte iron recicling, not at all. So maybe am I doing something wrong?
I followed the standard Perl's reaction:
- samples to water
- potassium ferrocyanide 2 % + Hidrochloric acid 2% at equal parts for 5, 10 and 30 minutes
- distilled water
- neutral red
-distilled water
-dehydrate and mount
thank you so much!
Hi Oscar. I don't know in fish, but in mice we can see very clear the blue in the spleen (red pulp) but almost nothing in the liver. Same as you. I can see in papers that the load of iron in the liver is much more lower than the spleen. Maybe, although the liver is a place for iron metabolism, does not accumulate/storage iron like in the spleen? I hope some expert can answer your question!
The protocol is used to identify ferric iron, such as in hemosiderin and ferritin. Not the ferrous form that is found in hemoglobin or myoglobin or nadph oxidase. Hemosiderin is found normally in tissues that are involved in rbc recycling likespleen and in hematopoiesis like red bone marrow, as well in recovered bleeds. Typically not in the liver, except in conditions of iron overload such as in hemachromatosis. Perhaps also if there is extramefullsrly hematopoiesis going on in the liver, but this is not really common.
Hello Lora and thanks for your answer!!
Indeed I found this papaer that shows it https://www.scielo.br/pdf/ni/v15n1/1982-0224-ni-15-01-e160041.pdf
so i feel very lost now.
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