Immunohistochemical assays in prostatic biopsies processed in Bouin's fixative.
OBJECTIVE
To investigate the problems involved in undertaking immunohistochemistry (IHC) and nuclear morphometry using Bouin's fixed prostate biopsies.


METHODS
Archival Bouin's fixed and formalin fixed, paraffin wax embedded prostatic biopsies were immunostained for three nuclear biomarkers (minichromosome maintenance protein 2 (MCM-2), p27, and Ki-67), one membrane localised biomarker (C-erb-B2), CD34, and alpha methylacyl-CoA racemase (AMACR). The quality of IHC staining was compared between tissues prepared separately in both fixatives. Feulgen staining was also performed on Bouin's fixed tissues to check its suitability for nuclear morphometry.


RESULTS
MCM-2 staining was completely negative in Bouin's fixed tissues, whereas p27 showed more background and excess cytoplasmic staining in Bouin's fixed versus formalin fixed tissues. C-erb-B2 showed non-specific, strong luminal cell staining in the Bouin's fixed tissue. Feulgen staining was also very weak in Bouin's fixed tissue. However, Ki-67, AMACR, and CD34 worked equally well in Bouin's and formalin fixed tissues.


CONCLUSIONS
Bouin's fixed tissues may be unsuitable when subsequent IHC and morphometry are contemplated. An awareness of which antibodies are suitable for use in Bouin's fixed biopsies is essential.
Ananthanarayanan
V
V
The Department of Preventive Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. viju@northwestern.edu
Pins
M R
MR
Meyer
R E
RE
Gann
P H
PH
D019342:Acetic Acid; D014408:Biomarkers, Tumor / Q000032:analysis*; D001706:Biopsy; D018797:Cell Cycle Proteins / Q000032:analysis; D002467:Cell Nucleus / Q000737:chemistry; D050760:Cyclin-Dependent Kinase Inhibitor p27; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D008297:Male; D064111:Minichromosome Maintenance Complex Component 2; D009363:Neoplasm Proteins / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D016612:Paraffin Embedding; D010851:Picrates; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000473:pathology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis; D012394:Rosaniline Dyes; D016707:Tissue Fixation / Q000379:methods*; D025521:Tumor Suppressor Proteins / Q000032:analysis
D019342:Acetic Acid; D014408:Biomarkers, Tumor; D018797:Cell Cycle Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27; D005404:Fixatives; D005557:Formaldehyde; D064111:Minichromosome Maintenance Complex Component 2; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D010851:Picrates; D018719:Erb-b2 Receptor Tyrosine Kinases; D012394:Rosaniline Dyes; D025521:Tumor Suppressor Proteins; C007722:Feulgen stain; C026239:Bouin's solution
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