Jul 05, 2023
Tissue biopsy FFPE preparation
- 1Johns Hopkins University
Protocol Citation: Kyu Sang Han, Pei-Hsun Wu 2023. Tissue biopsy FFPE preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzx81rgx1/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: July 03, 2023
Last Modified: July 05, 2023
Protocol Integer ID: 84426
Abstract
Creating human tissue FFPE block from excisional biopsy, and sectioning to create H&E slides.
Tissue biopsy collection
Tissue biopsy collection
Take excisional biopsy of human skin in abdomen or scalp
After excisional biopsy from a operating room (OR), place the skin tissue into a tissue container prefilled with buffered formalin for 12-24 hours at room temperature (RT)
Discard formalin, rinse with PBS, refill PBS, and leave the tissue in PBS for 1 minute
Take tissue out on a large Petri dish and measure the tissue size
Label a slotted cassette with a pencil with tissue ID and place the tissue in the cassette
Put cassette with the tissue back into the tissue container filled with PBS
Paraffin Embedding
Paraffin Embedding
Dehydrate tissue by submerging cassettes into ethanol at increasing concentrations from 70% to 99%.
Embed cassette paraffin at 60 deg C and store at room temperature
FFPE sectioning
FFPE sectioning
Section tissue and place on water bath to expand
Place expanded tissue section on a Superfrost Plus microscope slide
H&E stain
H&E stain
Stain with Hematoxylin and Eosin
Place coverslip