Nov 14, 2023

Public workspaceTissue Sectioning

DOI
dx.doi.org/10.17504/protocols.io.bp2l6xm8klqe/v1
  • Michael X. Henderson1
  • 1Van Andel Institute
Michael Henderson
Open access
DOI: dx.doi.org/10.17504/protocols.io.bp2l6xm8klqe/v1
Protocol Citation: Michael X. Henderson 2023. Tissue Sectioning. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6xm8klqe/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: November 06, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 90648
Keywords: ASAPCRN
Abstract
This protocol details formalin-fixed paraffin-embedded tissue sectioning on a microtome for subsequent tissue staining.
Attachments
n2fcbpqrp.pdf
n2fcbpqrp.pdf
161KB
Tissue Sectioning
Tissue Sectioning
2h 20m
Clean microtome blade with xylene (to remove paraffin) and then 100% ethanol (to dry).
Mount blade on microtome, and tighten screws to hold it in place.
Mount paraffin block in holder. Manually move the block near, but not touching the blade.
Align the block to the blade in both left-right and top-bottom directions by iterative adjustments
followed by checking the alignment.
Once the block is directly apposed to the blade and aligned, switch cutting depth to Thikness6 µm .

Hydrate block if needed with ammonia. Dry block after hydration.
Begin sectioning the block. Grasp the first section lightly. Continue sectioning until you have a
ribbon of sections (~6-10 sections).
Use camel hair brush to lightly remove ribbon by brushing from one side to the other. Alternatively, lightly separate the section attached to the block from the remainder of the sections by scoring with a razor blade.
Note
This will allow you to keep a non-wrinkled section attached to the front of the next ribbon.

Transfer the ribbon to a Temperature37 °C water bath. Burst bubbles with a syringe needle. Score between
sections with a razor blade.
Gently spring apart sections with forceps and mount them in order on glass slides. Transfer the
slides to an upright rack to allow the water to wick off of the sections before transferring to a
metal rack.
Continue sectioning (Steps 6-10) until the block is depleted of tissue.
Transfer slides to oven to dry at Temperature42 °C for Duration02:00:00 or more or Temperature60 °C for Duration00:20:00 or more.

2h 20m
Overnight