Dec 10, 2024
KAPP-Sen TMC: Tissue Sectioning Guidelines (FFPE) - CODEX/PhenoCycler
- 1The Jackson Laboratory
- Cellular Senescence Network (SenNet) Method Development Community
Protocol Citation: Santhosh Sivajothi, Emily Soja, William F Flynn, Elise Courtois 2024. KAPP-Sen TMC: Tissue Sectioning Guidelines (FFPE) - CODEX/PhenoCycler. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn96bzl5d/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: December 10, 2024
Last Modified: December 10, 2024
Protocol Integer ID: 114627
Keywords: CODEX, CODEX slide prep, tissue sectioning CODEX, KAPP-Sen TMC
Funders Acknowledgements:
National Institute on Aging (NIA) KAPP-Sen Tissue Mapping Collaborative
Grant ID: U54 AG075941
Abstract
The purpose of this SOP is to provide general guidelines for preparing FFPE tissue sections suitable for PhenoCycler/CODEX
Materials
Standard 1” x 3” positively charged glass microscopy slides (example, SuperFrost Plus)
Tissue Sectioning Guidelines for CODEX/PhenoCycler
Tissue Sectioning Guidelines for CODEX/PhenoCycler
Prepare the microtome for sectioning. Section the tissue at a thickness of 5 μm.
Note
Do not exceed 10 μm as this will affect the autofocusing capabilities of the microscope. Avoid folds and tears in the tissue, as these artifacts will affect image quality and data analysis.
For best results, the tissue should be completely adhered to the slide with minimal tears or folds.
Gently place the tissue section in the center of the slide within the imageable area as shown in
grey rectangle below.
If tissue overlaps the boundary, proper coverslip seal will not be formed. Multiple tissues can be placed within the boundary, as long as there is no overlap of paraffin over adjacent tissues
Working area dimensions on slide for optimal tissue placement
Example of good tissue placement:
Slide with CODEX coverslip showing optimal placement of tissue section
Dry tissue sections overnight at RT
After drying, store cut slides at 4°C (for up to six months)
NOTE: Do not place any stickers or labels on the frosted end of the slide as it may interfere with coverslip adhesion. Use markers or directly print labels on the slide.