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Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Immunohistochemical assays in prostatic biopsies processed in Bouin's fixative
V Ananthanarayanan1, M R Pins, R E Meyer
1The Department of Preventive Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. viju@northwestern.edu
Journal of Clinical Pathology
|March 1, 2005
Summary
Bouin's fixation is often unsuitable for prostate biopsies undergoing immunohistochemistry (IHC) and nuclear morphometry. While some biomarkers like Ki-67 work well, others like MCM-2 show negative staining, impacting diagnostic accuracy.
Area of Science:
- Pathology
- Biomarker analysis
- Histology
Background:
- Immunohistochemistry (IHC) and nuclear morphometry are crucial techniques in prostate cancer diagnosis.
- The choice of tissue fixative significantly impacts the quality of IHC staining and morphometric analysis.
- Bouin's fixative is an alternative to formalin, but its suitability for specific biomarkers needs evaluation.
Purpose of the Study:
- To assess the challenges and suitability of using Bouin's fixed prostate biopsies for immunohistochemistry (IHC) and nuclear morphometry.
- To compare the efficacy of IHC staining for various biomarkers in Bouin's fixed versus formalin-fixed tissues.
- To evaluate Feulgen staining in Bouin's fixed tissues for nuclear morphometry.
Main Methods:
- Archival prostate biopsies fixed in Bouin's solution and formalin were used.
- Immunohistochemical staining was performed for nuclear (MCM-2, p27, Ki-67), membrane (C-erb-B2), and other markers (CD34, AMACR).
- Feulgen staining was conducted on Bouin's fixed tissues to assess suitability for nuclear morphometry.
Main Results:
- MCM-2 staining was negative in Bouin's fixed tissues.
- p27 showed increased background and cytoplasmic staining, while C-erb-B2 exhibited non-specific luminal staining with Bouin's fixation.
- Feulgen staining was weak in Bouin's fixed tissues; however, Ki-67, AMACR, and CD34 performed comparably in both fixatives.
Conclusions:
- Bouin's fixed prostate biopsies may not be suitable for subsequent IHC and nuclear morphometry.
- Awareness of antibody suitability for Bouin's fixed tissues is essential for accurate diagnostic interpretation.
- Formalin fixation remains a more reliable choice for a broader range of IHC and morphometric analyses in prostate biopsies.
