Related Experiment Video
Updated: Jun 4, 2026
10:49
Automated Multiplex Immunofluorescence Panel for Immuno-oncology Studies on Formalin-fixed Carcinoma Tissue Specimens
Published on: January 21, 2019
Multiple immunofluorescence labeling of formalin-fixed paraffin-embedded tissue
David Robertson1, Clare M Isacke
1Breakthrough Centre, The Institute of Cancer Research, Royal Cancer Hospital, London, UK.
Methods in Molecular Biology (Clifton, N.J.)
|March 4, 2011
Summary
This study presents an optimized method for multiple immunofluorescent labeling of formalin-fixed paraffin-embedded (FFPE) tissues. This technique overcomes autofluorescence issues, enabling detailed analysis of archival tissue samples.
Area of Science:
- Histology
- Immunofluorescence
- Microscopy
Background:
- Multiple immunofluorescent labeling of FFPE tissue is not routine.
- Autofluorescence of FFPE tissue is a major limitation.
- Immunohistochemistry or cryosectioning are common alternatives.
Purpose of the Study:
- To describe an optimized method for high-resolution immunofluorescence labeling of FFPE tissue.
- To overcome FFPE autofluorescence for immunofluorescence applications.
- To enable multi-labeling investigations on archival FFPE samples.
Main Methods:
- Optimized protocol for FFPE tissue preparation.
- Combination of antigen retrieval and indirect immunofluorescence.
- Confocal laser scanning microscopy for imaging.
Main Results:
- Successful high-resolution immunofluorescence labeling of FFPE tissue achieved.
- Method overcomes inherent FFPE autofluorescence.
- Prepared samples are stable for long-term storage (>250 days) at -20°C with minimal quality loss.
Conclusions:
- This optimized method allows for robust multiple immunofluorescent labeling of FFPE tissues.
- The technique facilitates the use of large archival FFPE sample collections for immunofluorescence studies.
- This approach expands the utility of FFPE samples in biomedical research.
