Overview
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
- 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).
- Fill a 1.5 mL single-use polypropylene tube with 80 µL of digestion/lysis buffer (see Table of Materials).
- Put a clean pipette tip into the buffer.
- Identify cancer tissues of the tumor area most suitable for macrodissection according to the appropriate H&E-stained section.
- Macrodissect cancer tissue based on the marked H&E-stained section.
- Take a clean razor blade and gently scrape the cancer tissue off the slide, trying to keep it in one piece so that it is easier to work with.
- Take the pipette tip out of a single-use polypropylene tube containing buffer and use it to transfer the scrapped tissue into the buffer vial (see Table of Materials).
NOTE: The wet tip should attract the tissue, which makes the transfer easier.
- Repeat step 1.5 with the rest of the slides.
- After all the tissue is in the single-use polypropylene tube, use the tip to make sure the tissue is fully submerged and is not stuck to the wall of the tube.
- Add 20 µL of a subtilisin-related serine protease to the vial and gently flick to mix (see Table of Materials).
- Place the vial in a 55 °C heat block for at least 4 h or overnight. Make sure to slightly vortex after 2 h.
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Materials
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. |