I haven't had any experience regarding pathological examination. In order to examine the embryo of histopathological, I would like to stain by getting paraffin. However, I can't manage to do. When paraffin is taken, the samples are spoilt. I would really be grateful if you could explain me a method that I can easily figure out.
So thanks....
I agree with the previous comment. It sounds like you have not fixed your tissues first. For paraffin embedding and basic staining (e.g, H&E), a typical fixative is 10% neutral buffered formalin (available from most big suppliers).
Here are some important factors that influence how your specimen fixes: http://pathology.wustl.edu/research/cores/amp/index.php?page=Formalin%20Fixation
Fixation time will vary depending on the size of your specimen. Small specimens will fix overnight (at room temperature), but if it is large and/or the tissue is thick, then it can take up to 48-72 hours. If the fixative does not penetrate even after all of that time then consider using a fine tip pipette to deliver fixative into hard-to-reach places (e.g., mouth) and/or making some small incisions in the thickest parts of the animal before fixing.
Once it is fixed, you can place it in vessel of your choosing (agarose, tissue cassettes, etc.) and take it through the ethanol, clearing, embedding, sectioning, and staining steps.
http://library.med.utah.edu/WebPath/HISTHTML/HISTOTCH/HISTOTCH.html
https://www.researchgate.net/post/4_paraformaldehyde_for_fixing_cells-4_degrees_or_room_temp
You need to first fix the tissues and dehydrate them before you float in the paraffin.
Here is a link with a protocol: http://protocolsonline.com/histology/sample-preparation/paraffin-processing-of-tissue/
http://protocolsonline.com/histology/sample-preparation/paraffin-processing-of-tissue/
I agree with the previous comment. It sounds like you have not fixed your tissues first. For paraffin embedding and basic staining (e.g, H&E), a typical fixative is 10% neutral buffered formalin (available from most big suppliers).
Here are some important factors that influence how your specimen fixes: http://pathology.wustl.edu/research/cores/amp/index.php?page=Formalin%20Fixation
Fixation time will vary depending on the size of your specimen. Small specimens will fix overnight (at room temperature), but if it is large and/or the tissue is thick, then it can take up to 48-72 hours. If the fixative does not penetrate even after all of that time then consider using a fine tip pipette to deliver fixative into hard-to-reach places (e.g., mouth) and/or making some small incisions in the thickest parts of the animal before fixing.
Once it is fixed, you can place it in vessel of your choosing (agarose, tissue cassettes, etc.) and take it through the ethanol, clearing, embedding, sectioning, and staining steps.
http://library.med.utah.edu/WebPath/HISTHTML/HISTOTCH/HISTOTCH.html
https://www.researchgate.net/post/4_paraformaldehyde_for_fixing_cells-4_degrees_or_room_temp
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