Feb 13, 2026

Public workspaceH&E and Immunohistochemistry (IHC) Staining

DOI
https://dx.doi.org/10.17504/protocols.io.n2bvj1j6wvk5/v1
  • Guangli Suo1
  • 1CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Jiangsu, 215123, China.
  • MTS
Guangli Suo
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DOI: https://dx.doi.org/10.17504/protocols.io.n2bvj1j6wvk5/v1
Protocol Citation: Guangli Suo 2026. H&E and Immunohistochemistry (IHC) Staining. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvj1j6wvk5/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 13, 2026
Last Modified: February 13, 2026
Protocol Integer ID: 243187
Keywords: immunohistochemistry, brc tissue, ihc
Abstract
H&E and Immunohistochemistry (IHC) Staining for MTS and BRC tissue.
Materials
- MWC-chips
- PBS
- HACS technology
- Centrifuge
- Agar
- Paraformaldehyde
- Ethanol
- Citrate Unmasking Solution (Beyotime)
- Antibodies against ER, PR, HER2, Ki-67 (Proteintech)
- BSA solution
- Diaminobenzidine (DAB) (Solarbio)
- NanoZoomer S210 microscope (Visiopharm, Denmark)
Troubleshooting
H&E and Immunohistochemistry (IHC) Staining
Wash MTSs from MWC-chips with PBS and sort using HACS technology.
Collect MTSs through centrifugation at 1,200 rpm and 4 °C for 10 minutes.
Wash with cold PBS and embed MTSs in agar.
Fix agar-embedded MTSs and primary tumor tissues overnight in PBS with 4% paraformaldehyde.
Perform paraffin-embedding, sectioning, and H&E staining.
Dewax and rehydrate paraffin-embedded MTSs or BRC tissue sections through a graded ethanol series.
Perform heat-mediated antigen retrieval with Citrate Unmasking Solution.
Block sections at room temperature for 1 hour.
Perform IHC staining overnight at 4 °C with antibodies against ER, PR, HER2, Ki-67.
Dilute antibodies in a ratio of 1:200 in 3% BSA solution.
Use DAB to achieve optimal staining intensity.
Visualize stained tissues using the NanoZoomer S210 microscope.