Paraffin Embedding of Tissue Samples: Difference between revisions
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==SOP== | |||
*[[SOP - Carcinogens]] | |||
*[[SOP - Flammables and Combustibles]] | |||
*[[SOP - Formalin and Paraformaldehyde]] | |||
*[[SOP- Irritants]] | |||
== Paraffin embedding protocol == | == Paraffin embedding protocol == | ||
#Harvest tissue from animal, place into cassette and submerge in 10% neutral buffered formalin for 24-48 hrs (depending on thickness of sample). | #Harvest tissue from animal, place into cassette and submerge in 10% neutral buffered formalin for 24-48 hrs (depending on thickness of sample). | ||
#Move cassettes into 70% ethanol for 24 hrs. Samples can be left in 70% for long term storage, if required. | #Move cassettes into 70% ethanol for 24 hrs. Samples can be left in 70% for long term storage, if required. At this step samples can also be taken to the [https://www.rogelcancercenter.org/research/shared-resources-and-cores/tissue-and-molecular-pathology Rogel Cancer Center's Tissue and Molecular Pathology Core] for the remaining steps | ||
##Note that the subsequent dehydration steps involve many long incubation periods and therefore should be started first thing in the morning (unless you plan on being in the lab overnight!). | ##Note that the subsequent dehydration steps involve many long incubation periods and therefore should be started first thing in the morning (unless you plan on being in the lab overnight!). | ||
#Turn on wax pots. | #Turn on wax pots. | ||
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#Store at room temperature until ready to section. | #Store at room temperature until ready to section. | ||
Wash Solution Table | ==Wash Solution Table== | ||
{|style="border-collapse: separate; border-spacing: 0; border-width: 1px; border-style: solid; border-color: #000; padding: 0" | {|style="border-collapse: separate; border-spacing: 0; border-width: 1px; border-style: solid; border-color: #000; padding: 0" | ||
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|style="border-style: solid; border-width: 0 1px 0 0"| 95% ethanol | |style="border-style: solid; border-width: 0 1px 0 0"| 95% ethanol | ||
|style="border-style: solid; border-width: 0"| | |style="border-style: solid; border-width: 0"| 75 min | ||
|- | |||
|style="border-style: solid; border-width: 0 1px 0 0"| 95% ethanol | |||
|style="border-style: solid; border-width: 0"| 75 min | |||
|- | |||
|style="border-style: solid; border-width: 0 1px 0 0"| 100% ethanol | |||
|style="border-style: solid; border-width: 0"| 60 min | |||
|- | |||
|style="border-style: solid; border-width: 0 1px 0 0"| 100% ethanol | |||
|style="border-style: solid; border-width: 0"| 60 min | |||
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|style="border-style: solid; border-width: 0 1px 0 0"| 100% ethanol | |style="border-style: solid; border-width: 0 1px 0 0"| 100% ethanol | ||
|style="border-style: solid; border-width: 0"| | |style="border-style: solid; border-width: 0"| 60 min | ||
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|style="border-style: solid; border-width: 0 1px 0 0"| Citrosolv | |style="border-style: solid; border-width: 0 1px 0 0"| Citrosolv | ||
|style="border-style: solid; border-width: 0"| | |style="border-style: solid; border-width: 0"| 30 min | ||
|- | |||
|style="border-style: solid; border-width: 0 1px 0 0"| Citrosolv | |||
|style="border-style: solid; border-width: 0"| 30 min | |||
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|style="border-style: solid; border-width: 0 1px 0 0"| 60 degree paraffin | |||
|style="border-style: solid; border-width: 0"| 60 min | |||
|- | |||
|style="border-style: solid; border-width: 0 1px 0 0"| 60 degree paraffin | |||
|style="border-style: solid; border-width: 0"| 60 min | |||
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|style="border-style: solid; border-width: 0 1px 0 0"| 60 degree paraffin | |style="border-style: solid; border-width: 0 1px 0 0"| 60 degree paraffin | ||
|style="border-style: solid; border-width: 0"| | |style="border-style: solid; border-width: 0"| 60 min or O/N | ||
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