FFPE tissue shows no staining from incomplete deparaffinization

Symptom

Paraffin-embedded tissue sections show no immunostaining. Residual paraffin creates a physical barrier preventing antibody access to tissue antigens.

1. 1 Insufficient deparaffinization time leaving paraffin residue in tissue
2. 2 Old or degraded xylene with reduced solvent effectiveness
3. 3 Incomplete paraffin removal blocking antibody penetration into tissue sections

1. 1 Deparaffinize sections for longer duration
2. 2 Use fresh xylene for optimal paraffin removal
3. 3 Perform additional xylene incubation steps (3 changes of 5 min each)
4. 4 Verify complete deparaffinization by checking tissue appearance before proceeding

Related Video (3)

Cell Signaling Technology ★ 90

Immunohistochemistry Protocol for Paraffin embedded Tissue Sections

"Full IHC protocol on paraffin sections includes the deparaffinization steps where failure occurs."

Bilibili (China-Accessible Mirrors) ★ 85

Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)

"Step-by-step demo on FFPE sections covers deparaffinization and antigen retrieval, directly addressing the barrier issue."

Bilibili (China-Accessible Mirrors) ★ 75

Paraffin Section IHC Experimental Workflow

"Workflow video for paraffin sections shows the deparaffinization stage that likely caused the no-staining problem."