Rapid microwave-stimulated fixation of entire prostatectomy specimens. Biomed-II MPC Study Group.
Author(s): Ruijter ET, Miller GJ, Aalders TW, van de Kaa CA, Schalken JA, Debruyne FM, Boon ME
Publication: J Pathol, 1997, Vol. 183, Page 369-75
PubMed ID: 9422995 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of method of fixative delivery on the histology of prostate specimens obtained at autopsy.
Summary of Findings:
Sectioning of specimens which were fixed through immersion in neutral buffered formalin or microwave-accelerated fixation for 6 min followed by 3.5 min was easy, regardless of whether the tissue was previously perfused through multiple injections of formalin. However, if the tissue underwent microwave-accelerated fixation for only 6 min, with or without perfusion, sectioning was difficult and resulted in damage to the tissue. Tissue that was not perfused with formalin prior to H&E staining displayed an edge effect. The authors report that there were no significant differences between conventional immersion and microwave-accelerated fixation for PAS, van Gieson's elastica or Alcian blue staining.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform Morphology H-and-E microscopy Morphology Light microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Method of fixative delivery Immersion
Microwaved
Injection
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Study Purpose
The purpose of this study was to determine the effects of method of fixative delivery on immunohistochemistry (IHC) of prostate specimens obtained at autopsy.
Summary of Findings:
E-cadherin staining required antigen retrieval and was most intense when conventional immersion fixation was for 8-12 h. When conventional immersion fixation was used, most E-cadherin staining was peripheral or absent with only 16% of specimens displaying specific homogeneous staining. Multiple perfusion injections prior to immersion fixation increased the number of specimens with specific homogeneous E-cadherin staining to 53%. When perfusion was followed by two rounds of microwave irradiation, 93% of the specimens had specific homogeneous E-cadherin staining. Method of fixative delivery and duration of fixation had no effects on p53 or MIB-1 staining. Cytokeratin staining in conventionally fixed specimens decreased slightly when fixation was for more than 12 h, but was increased by protease digestion. Cytokeratin staining was homogeneous in all specimens fixed by perfusion followed by two rounds of microwave irradiation.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Method of fixative delivery Immersion
Injection
Microwaved
Biospecimen Preservation Time in fixative 1 h
2 h
3 h
4 h
8 h
12 h
18 h
24 h
36 h
Immunohistochemistry Specific Targeted peptide/protein Cytokeratin
E-cadherin
MIB-1
p53
Analyte Extraction and Purification Antigen retrieval Microwave in 0.1 M citrate buffer
None
Analyte Extraction and Purification Protein digestion 10 min digestion in Protease XIV at 37 degrees C
None
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Study Purpose
The purpose of this study was to determine the effects of method of fixative delivery on the size of extracted DNA from prostate specimens obtained at autopsy.
Summary of Findings:
When specimens were fixed by immersion in formalin for 20 h, only 34% of the DNA was over 1 kb and 2-3% was over 4 kb. In contrast, when the method of fixative delivery included multiple perfusion injections followed by two rounds of microwave irradiation, 78% of the extracted DNA was over 1 kb and 40% was over 4 kb.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform DNA Electrophoresis Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Method of fixative delivery Immersion
Microwaved
Injection