Knowledge Base › Troubleshooting Atlas › IHC
Cytoplasmic bleed with a nuclear marker
What it looks like
Cytoplasmic staining appears diffuse or irregularly distributed alongside the nuclear marker, obscuring nuclear details.
Where it usually appears: Commonly observed in poorly fixed tissues or during prolonged antigen retrieval.
Likely causes
- Inadequate fixation
- Overheating during antigen retrieval
- Prolonged incubation with primary antibody
- Non-specific binding of antibodies
Immediate checks
- Check fixation time and method used
- Review antigen retrieval protocol
- Inspect primary antibody concentration
Corrective action
- Re-fix the tissue with appropriate fixative
- Optimize antigen retrieval conditions
- Dilute primary antibody
Preventive action
- Ensure proper fixation and processing of tissues
- Standardize antigen retrieval protocols
- Use high-quality, specific antibodies
This atlas is for educational purposes for laboratory professionals. It does not replace your institutional SOPs, manufacturer instructions, validated protocols or pathologist judgment. Adapt any action to your laboratory and equipment.
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