Dec 09, 2024

Public workspaceParaffin section tissue preparation

DOI
dx.doi.org/10.17504/protocols.io.6qpvr9353vmk/v1
  • Ashley Seifert1
  • 1University of Kentucky
Ashley Seifert
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DOI: dx.doi.org/10.17504/protocols.io.6qpvr9353vmk/v1
Protocol Citation: Ashley Seifert 2024. Paraffin section tissue preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr9353vmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 06, 2024
Last Modified: December 09, 2024
Protocol Integer ID: 114534
Keywords: ASAPCRN
Disclaimer
Note that any protocol involving animals should be reviewed and approved by your Institutional Animal Care and Use Committee (IACUC) before use.
Abstract
This protocol describes the procedure used by the Seifert Lab to prepare paraffin-embedded tissue sections.
Protocol materials
ReagentEppendorf tubes 1.5 mL uncoloredEppendorfCatalog #022363204
ReagentEpredia 10% Neutral Buffered FormalinThermo Fisher ScientificCatalog #5701
Reagent1X PBS (Phosphate-buffered saline )
Reagent70% Ethanol
ReagentFormula R paraffinLeica BiosystemsCatalog #3801450
For each sample, prepare a ReagentEppendorf tubes 1.5 mL uncoloredEppendorfCatalog #022363204 with ReagentEpredia 10% Neutral Buffered FormalinThermo Fisher ScientificCatalog #5701

Harvest tissue from animal and immerse in 10% NBF in prepared tube
Incubate DurationOvernight at Temperature4 °C on a rocker (16-18 hr fix)

Remove the 10% NBF
Fill tube with Reagent1X PBS (Phosphate-buffered saline )

Wash at Temperature4 °C on rocker for Duration00:10:00

Drain and repeat steps 5 and 6 two more times for a total of 3 washes
Drain PBS and add Reagent70% Ethanol Contributed by users

Wash at Temperature4 °C on rocker for Duration00:10:00
Drain and repeat two more times for a total of 3 washes
After third wash, drain and replace with fresh 70% Ethanol
Store at Temperature4 °C until ready for processing

Process for ReagentFormula R paraffinLeica BiosystemsCatalog #3801450 embedding using a
Equipment
Histos 5
NAME
Tissue processor
TYPE
Milestone
BRAND
61577
SKU
https://www.milestonemedsrl.com/us/products/tissue-processing/?_gl=1*18i3dwh*_up*MQ..*_gs*MQ..&gclid=EAIaIQobChMIke623eqTigMVtszCBB3u3RJ2EAAYASAAEgJ_p_D_BwE&gclsrc=aw.ds
LINK


Section samples at Thikness5 µm