Feb 17, 2025

Public workspacePost MALDI IMS CODEX Staining and Imaging Fresh Frozen Tissue

DOI
dx.doi.org/10.17504/protocols.io.x54v9rb24v3e/v1
  • 1Vanderbilt University;
  • 2Delft University of Technology;
  • 3Division of Nephrology, Vanderbilt University Medical Center
  • VU Biomolecular Multimodal Imaging Center / Spraggins Research Group
    Tech. support email: jeff.spraggins@vanderbilte.du
Thai Pham
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DOI: dx.doi.org/10.17504/protocols.io.x54v9rb24v3e/v1
Protocol Citation: Thai Pham, Lukasz Migas, Madeline E. Colley, Ellie Pringy, Anna Smith, Jamie Allen, Mark De Caestecker, Melissa Farrow, Raf Van De Plas, Jeff Spraggins, Martin Dufrense 2025. Post MALDI IMS CODEX Staining and Imaging Fresh Frozen Tissue. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9rb24v3e/v1
Manuscript citation:
PhenoCycler-Fusion User Guide (Version 2.1.0) 2023 by Akoya Biosciences, Inc.
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, 2025
Last Modified: February 17, 2025
Protocol Integer ID: 120570
Abstract
PhenoCycler User Guide for multiplex spatial proteomic detection on post IMS Fresh Frozen tissue sections
Pre-Experiment Preparation
Pre-Experiment Preparation
Download PhenoCycler-Fusion User Guide_2.1.0.pdfPhenoCycler-Fusion User Guide_2.1.0.pdf
Prepare reagent and solutions as listed in the PhenoCycler-Fusion User Guide (Version 2.1.0) from page 34 to 36
Post IMS- CODEX FF Tissue Staining
Post IMS- CODEX FF Tissue Staining
Remove IMS matrix by gradually spraying acetone on the tissue slide, until all the matrix is washed out.
Submerge the slide in acetone in 2 minutes for 5 times.

Remove sample from acetone, let it sit in the staining box 2 minutes.
Download PhenoCycler-Fusion User Guide_2.1.0.pdfPhenoCycler-Fusion User Guide_2.1.0.pdf
Continue the procedure from Step 5 (page 38-44) of the PhenoCycler-Fusion User Guide (Version 2.1.0).
CODEX FF Tissue Imaging
CODEX FF Tissue Imaging
Download PhenoCycler-Fusion User Guide_2.1.0.pdfPhenoCycler-Fusion User Guide_2.1.0.pdf
Apply the flowcell onto tissue slide (page 29-39)
Instrument set-up for PhenoCycler scan (page 57-71)