Jan 03, 2025
Yale SenNet Human TMC - H&E staining following Phenocycler-Fusion Imaging
- Archibald Enninful1,
- Negin Farzad1,
- Jungmin Nam1,
- Yale Pathology Tissue Services1,
- Rong Fan1
- 1Yale University
- Fan Lab Yale
Protocol Citation: Archibald Enninful, Negin Farzad, Jungmin Nam, Yale Pathology Tissue Services, Rong Fan 2025. Yale SenNet Human TMC - H&E staining following Phenocycler-Fusion Imaging. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl8d6j9g2w/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: January 03, 2025
Last Modified: January 03, 2025
Protocol Integer ID: 117628
Keywords: phenocycler-fusion, codex, h&e
Funders Acknowledgements:
Rong Fan
Grant ID: 1U54AG076043-01
Abstract
This protocol was adapted from (Yale Murine TMC - H&E staining following Phenocycler-Fusion Imaging)
Flow cell removal after Phenocycler-Fusion imaging and following H&E staining steps from Yale Pathology Tissue Services (YPTS). For FFPE samples, the tissue section is deparaffinized and rehydrated during the preprocessing steps for CODEX, hence these steps do not need to be repeated following theremoval of the flow cell
Removing flow cell following CODEX imaging
Removing flow cell following CODEX imaging
Remove slides from storage buffer.
Immerse slides to xylene or HistoClear overnight. However, an incubation of 20 minutes should suffice.
10m
Separate flow cell from the slide using a razorblades.
Rinse the tissue sample with DI water using at least 1L of DI water to ensure thorough rinsing.
Next, wash the tissue sample slide in 1x PhenoCyclerTM Buffer without additive, 3 x 10 minutes.
If you are not performing H&E immediately, transfer the tissue sample slide(s) to the PhenoCycler Storage
buffer for storage.
Hematoxylin and eosin staining
Hematoxylin and eosin staining
11m 30s
11m 30s
Stain with hematoxylin for 5 min.
5m
Rinse with water.
Clarifier for 30 seconds.
30s
Rinse with water.
Bluing for 1 min.
1m
Rinse with water.
Stain with eosin 5 min.
5m
Immerse slides in graded ethanol and clear slide in xylene.
Add coverslip with resin mounting medium.
Acknowledgements
Entire Yale SenNet TMC as well as the Yale Pathology Tissue Services