Dec 07, 2023

Public workspaceGeneral Microtome Sectioning of Formalin-Fixed Paraffin Embedded (FFPE) blocks

DOI
dx.doi.org/10.17504/protocols.io.bp2l69morlqe/v1
  • 1Department of Clinical and Movement Neurosciences, UCL Institute of Neurology, Queen Square, London WC1N 3BG, UK;
  • 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20 815, USA.;
  • 3QSBB, 1 Wakefield Street London Greater London WC1N 1PJ, UK
Toby J Curless
Open access
DOI: dx.doi.org/10.17504/protocols.io.bp2l69morlqe/v1
Protocol Citation: Toby J Curless, Zane Jaunmuktane 2023. General Microtome Sectioning of Formalin-Fixed Paraffin Embedded (FFPE) blocks. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l69morlqe/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: February 17, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 77172
Keywords: ASAPCRN, Tissue, Sectioning, Parkinson's disease, Microtome, Formalin-fixed Paraffin Embedded
Funders Acknowledgement:
The Michael J. Fox Foundation for Parkinson’s Research (MJFF) and the Aligning Science Across Parkinson’s (ASAP) Initiative
Grant ID: ASAP-000478
Abstract
This protocol describes the steps for the sectioning of formalin-fixed paraffin embedded blocks in preparation for further tissue processing such as immunohistochemistry.
Materials
SuperfrostTM charged microscope slides
Microtome
Fine tweezers
Fine paintbrush (optional)
Water bath
Slide holder
Safety warnings
Attention
Follow approved local risk assessment and institutional policies on the use of sharps.
Before start
  • Check all equipment, supplies and consumables
  • Set-up water bath to required temperature
  • Ensure blade is fit-for-purpose and clean
  • Label/barcode SuperFrost Plus slides
Preparation
Preparation
Check all equipment, consumables and supplies are in place.
Place a clean blade into the blade holder of the microtome.
Place the FFPE block onto ice, with the face of the block on the surface of the TemperatureOn ice until the block is cold through.

Heat Amount1 L distilled H2O toTemperature40 °C in a water bath.

Incubation
Sectioning
Sectioning
12h
Once both block and water are at the correct temperature, insert the block into the chuck of the microtome.
Adjust the levers on the microtome to position the face of the block parallel with the blade.


Lower the block until it is level with the blade and move the block closer to the blade using the handle on the microtome.
Select the thickness of choice on the microtome (usually Thikness4-7 µm .

Section and trim into the block until the whole tissue cross-section is present in the sections.
Using tweezers or a fine paintbrush, gently pull sections down until a ribbon of tissue sections has formed.

Pick up the ribbon with tweezers and place into the water bath to allow wrinkles to disappear from the tissue.
A ribbon of four consecutive sections floating in the water bath to remove wrinkles.

Separate sections using tweezers and collect individual sections on SuperFrostTM charged microscope slides.
Allow slides to drain and air dry vertically at TemperatureRoom temperature for > Duration12:00:00 prior to further processing.

12h
Critical