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Immunohistochemistry (No Staining) moderate

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.
Common Causes
  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
Solutions
  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)
Bilibili (China-Accessible Mirrors) ★ 92
Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)
"Directly demonstrates IHC on FFPE tissue with explicit coverage of antigen retrieval and deparaffinization steps, using CST protocol that addresses the root cause of incomplete deparaffinization"
Cell Signaling Technology ★ 85
Better IHC Step 1: Antigen Retrieval
"Focuses specifically on antigen retrieval optimization, which depends critically on proper deparaffinization; teaches troubleshooting for poor staining results caused by inadequate tissue preparation"
Cell Signaling Technology ★ 78
Immunohistochemistry Protocol for Paraffin embedded Tissue Sections
"Comprehensive IHC protocol video for FFPE tissue sections that covers the complete deparaffinization and tissue preparation workflow essential for preventing this failure mode"
Source: abcam.com ↗
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