FFPE Tumor Tissue Macrodissection: A Technique to Obtain Specific Tumorous Tissue from Unstained Specimens

Abstract

Source: Sugimoto, K. et al. Genome-Wide Analysis of DNA Methylation in Gastrointestinal Cancer. J. Vis. Exp. (2020)

This video describes a method for the manual macrodissection of cancer tissue from unstained tissue specimens to obtain a specific cancer tissue. The tissue section can be processed further to obtain DNA for genome-wide studies on gastrointestinal cancers.

Protocol

1. Scraping the Slides

1. Prepare digestion/lysis buffer with 650 mL of diethylpyrocarbonate (DEPC)-treated water, 100 mL of ethylenediaminetetraacetic acid (EDTA), 50 ml of Tris hydrochloride (Tris-HCL) 2 M pH 8.8, and 200 mL of 10% sodium dodecyl sulfate (SDS) (see Table of Materials).
2. Fill a 1.5 mL single-use polypropylene tube with 80 µL of digestion/lysis buffer (see Table of Materials).
3. Put a clean pipette tip into the buffer.
4. Identify cancer tissues of the tumor area most suitable for macrodissection according to the appropriate H&E-stained section.
5. Macrod....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
10% Sodium dodecyl sulfate (SDS)	Quality Biological	351-032-721 EA	Step 2.1.
DEPC-treated water	Quality Biological	351-068-131	Step 2.1.
Ethylenediaminetetraacetic acid (EDTA)	Corning	46-034-CL	Step 2.1.
Tris hydrochloride (Tris-HCL) 2 M pH 8.8	Quality Biological	351-092-101	Step 2.1.
Proteinase K	New England Biolabs	P8107S	Step 2.8.
Single-use polypropylene (Eppendorf) tube	Eppendorf	24533495	Step 2.5.2.