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

JoVE Encyclopedia of Experiments

Kankeronderzoek

0 weergaven • 2:31 min • April 30th, 2023

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

To begin macrodissection, take a glass slide carrying a tissue specimen. Additionally, take a reference slide carrying the same tissue specimen and stain it using hematoxylin and eosin for H&E stain.

In normal cells, hematoxylin imparts a blue stain to nucleic acids, while the eosin counterstain shows the cellular proteins pink. The deep blue coloration of the nucleus helps in differentiating cancer cells from normal cells.

Overlay the H&E stained slide with the unstained slide to align the tissue specimens. Locate the portion containing tumor cells on the H&E stained slide. Mark the corresponding area on the unstained slide.

Scrape off the desired tissue from the unstained slide. Using a wet pipette tip, collect the scraped tissue to ensure an easy transfer of the tissue specimen. Transfer the tissue into a tube containing a lysis buffer. Components of the lysis buffer digest the tissue, releasing the constituents of the cell.

Treat the lysate with a suitable protease and incubate at a high temperature. The protease cleaves the contaminating DNases, thereby protecting the DNA from degradation. Store the DNA containing tissue lysate for downstream analysis.

Identify cancer tissues of the tumor area most suitable for macrodissection according to the appropriate H&E stained section. Then, macrodissect the cancer tissue based on the marked section. Take a clean razor blade and gently scrape the cancer tissue off of the slide, trying to keep it in one piece. Use the pipette tip to transfer the scraped tissue into the lysis buffer vial.

Repeat this process with the rest of the slides. After all the tissue is in the tube, use the tip to make sure the tissue is fully submerged and is not stuck to the wall. Add 20 microliters of a subtilisin-related serine protease to the vial and gently flick to mix. Place the vial in a 55 degree Celsius heat block for at least 4 hours or overnight, making sure to slightly vortex after 2 hours.

07:37

SIVQ-LCM protocol voor de ArcturusXT Instrument

Gerelateerde video's

0 Bekeken

07:42

Laser-capture microdissectie van Human Prostatic Epithelium voor RNA-analyse

Gerelateerde video's

0 Bekeken

08:30

Bereiding van formaline gefixeerde paraffine-ingebed weefsel Cores voor zowel RNA en DNA-extractie

Gerelateerde video's

0 Bekeken

04:17

Formalin-Fixed Paraffin Embedded (FFPE) Tissue Preservation: A Method for Studying Tissue Morphology

Gerelateerde video's

0 Bekeken

03:53

Laser Microdissection: A Technique to Isolate Invading Colorectal Cancer cells from a 3D Organotypic Culture Model

Gerelateerde video's

0 Bekeken

09:20

Proteomics Monstervoorbereiding van formaline gefixeerde en in paraffine ingebed weefsel

Gerelateerde video's

0 Bekeken

10:33

Molecular Profiling van de invasieve tumor micro in een 3-dimensionaal model van colorectale kanker cellen en Ex vivo Fibroblasts

Gerelateerde video's

0 Bekeken

07:42

A Novel Microdissection aanpak voor het herstellen Mycobacterium tuberculosis Specifieke Afschriften van formaline gefixeerde in paraffine ingebedde Lung Granulomen

Gerelateerde video's

0 Bekeken

09:09

Laser Capture Microdissection of Glioma Subregions for Spatial and Molecular Characterization of Intratumoral Heterogeneity, Oncostreams, and Invasion

Gerelateerde video's

0 Bekeken

06:44

Geautomatiseerd dissectieprotocol voor tumorverrijking in weefsels met een laag tumorgehalte

Gerelateerde video's

0 Bekeken

Laatst bijgewerkt: 22 augustus 2026