Ultrasound-accelerated formalin fixation of tissue improves morphology, antigen and mRNA preservation
Author(s): Chu Wei-Sing, Furusato Bungo, Wong Kondi, Sesterhenn Isabell A, Mustofi Fathollah K, Wei Min Q, Zhu Zhenqing, Abbondanzo S, Liang Qi
Publication: Mod Pathol, 2005, Vol. 18, Page 850
PubMed ID: 15605077 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
To assess if tissue morphology is adequately preserved by USAFF (5-30 min) compared to conventional formalin fixation (16-24 h) using several tissues (brain, tonsil, lymph node, lung, prostate, soft tissue) obtained by biopsy or post mortem.
Summary of Findings:
Brain and lymphoid tissue fixed for 15-30 min by USAFF yielded tissue morphology comparable to conventional formalin fixation. Further, USAFF for 15 min resulted in enhanced eosin staining compared to other fixation durations and methods.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Not specified
- Autopsy
Platform:
Analyte Technology Platform Morphology H-and-E microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Time in fixative 15 min
30 min
16 h
Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
-
Study Purpose
To assess if protein antigenicity is adequately preserved by USAFF (5-30 min) compared to conventional formalin fixation (16-24 h) using several tissues (brain, tonsil, lymph node, lung, prostate, soft tissue) obtained by biopsy or post mortem.
Summary of Findings:
ER and CD50 immunoreactivity in breast and lymph node specimens, respectively, fixed for 5 min in the presence of ultrasound resulted in superior staining to conventional formalin fixation for 24 h; specifically, USAFF resulted in an increase in antigenicity and often eliminated the need for antigen retrieval. Further, a shortened staining method was successfully employed using USAFF tissue.
Biospecimens
- Tissue - Breast
- Tissue - Tonsil
- Tissue - Brain
- Tissue - Lymph Node
- Tissue - Lung
- Tissue - Prostate
- Tissue - Other
Preservative Types
- Formalin
Diagnoses:
- Not specified
- Autopsy
Platform:
Analyte Technology Platform Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
-
Study Purpose
To assess if RNA integrity is adequately preserved by USAFF (5-30 min) compared to conventional formalin fixation (16-24 h) using several tissues (brain, tonsil, lymph node, lung, prostate, soft tissue) obtained by biopsy or post mortem.
Summary of Findings:
Specimens conventionally fixed for 22 h displayed sporadic "edge effect" mRNA signals, while those fixed for 15 min in the presence of ultrasound displayed intense and location appropriate signals across the entire tissue section.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Not specified
- Autopsy
Platform:
Analyte Technology Platform RNA In situ hybridization Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
-
Study Purpose
To assess if protein abundance is adequately preserved by USAFF (5-30 min) compared to conventional formalin fixation (16-24 h) using several tissues (brain, tonsil, lymph node, lung, prostate, soft tissue) obtained by biopsy or post mortem.
Summary of Findings:
As determined by coomassie blue and Western blot analysis, a greater abundance of total and specific proteins, respectively, were observed in frozen and USAFF fixed tissue (5 min) than by conventional fixation (16 h).
Biospecimens
Preservative Types
- Formalin
- Frozen
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform Protein 1D/2D gels Protein Western blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
Snap frozen
Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
-
Study Purpose
To assess if mRNA abundance is adequately preserved by USAFF (5-30 min) compared to conventional formalin fixation (16-24 h) using several tissues (brain, tonsil, lymph node, lung, prostate, soft tissue) obtained by biopsy or post mortem.
Summary of Findings:
RNA integrity, as measured by RT-PCR analysis of beta-actin, was superiorly preserved in USAFF tissue compared to conventional fixation methods, with longer amplicons generated (548 bp) with USAFF than observed by conventional methods (158 bp sized product only).
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform RNA RT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) RT-PCR Specific Targeted nucleic acid beta-actin
RT-PCR Specific Length of gene fragment 548 bp
158 bp
Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated