Ultrasound-accelerated Tissue Fixation/Processing Achieves Superior Morphology and Macromolecule Integrity with Storage Stability
Author(s): Chu Wei-Sing, Liang Qi, Tang Yao, King Randy, Wong Kondi, Gong Maokai, Wei Minqi, Liu Jilan, Feng Shaw-Huey, Lo Shyh-Ching, Andriko Jo-Ann, Orr Marshall
Publication: J Histochem Cytochem, 2006, Vol. 54, Page 503
PubMed ID: 16314441 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
To determine if USAFF and processing compromises the stability of cellular morphology or protein antigenicity after prolonged storage of paraffin blocks at room temperature.
Summary of Findings:
No differences in cellular structure or preservation were observed in USAFF tissue as compared to conventional formalin fixed paraffin embedded tissue, as determine by microscopic analysis. Immunostaining revealed USAFF tissue yielded superior staining, required less primary antibody, and antigen retrieval was often unnecessary in comparison to conventional formalin fixation. Further, a shortened staining protocol could be employed with USAFF tissue. Storage of USAFF paraffin tissue blocks for up to 7 years did not affect immunohistochemical staining, indicating stability was preserved.
Biospecimens
- Tissue - Lung
- Tissue - Breast
- Tissue - Colorectal
- Tissue - Heart
- Tissue - Kidney
- Tissue - Brain
- Tissue - Liver
- Tissue - Lymph Node
- Tissue - Pancreas
- Tissue - Prostate
- Tissue - Skin
- Tissue - Spleen
- Tissue - Uterus
- Tissue - Blood Vessel
Preservative Types
- Formalin
Diagnoses:
- Autopsy
- Not specified
Platform:
Analyte Technology Platform Protein Immunohistochemistry Morphology H-and-E microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
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Study Purpose
To determine if USAFF and processing compromises RNA stability after prolonged storage of paraffin blocks at room temperature. In situ hybridization of kappa light chain immunoglobulin mRNA was compared among specimens freshly preserved by conventional formalin fixation and USAFF as well as USAFF tissue blocks after 7 years of storage at room temperature.
Summary of Findings:
Prolonged storage of USAFF paraffin blocks did not impact in situ hybridization results. Further, USAFF specimens stored for seven years displayed uniform and appropriate signal location, while signals in specimens freshly fixed by conventional methods were sporadic and limited to the periphery of the 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 Ultrasound-accelerated
Immersion
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Study Purpose
To determine if USAFF or long term storage of paraffin tissue blocks compromises the level of extracted DNA.
Summary of Findings:
Although greater DNA concentrations were observed in USAFF tissue compared to conventional formalin fixation, the abundance and success of PCR products were dependent on amplicon size, with larger amplicons requiring a higher cycle number and a complete lack of large (1 kb) products for all formalin fixed tissue.
Biospecimens
Preservative Types
- Formalin
- Frozen
Diagnoses:
- Not specified
- Autopsy
Platform:
Analyte Technology Platform DNA PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) PCR Specific Targeted nucleic acid beta-actin
PCR Specific Length of gene fragment 220 bp
489 bp
811 bp
1062 bp
647 bp
Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
Snap frozen
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Study Purpose
To determine if USAFF or long term storage of paraffin tissue blocks compromises RNA expression levels.
Summary of Findings:
Similar to DNA results, mRNA expression levels were dependent on the length of the generated amplicon, with a higher PCR success rate observed with shorter amplicons. After prolonged storage, conventionally fixed tissue and USAFF tissue yielded comparable results that were inferior to frozen tissue, as observed by semi-quantitative RT-PCR. However, real time quantitative RT-PCR analysis showed that a larger quantity of relative RNA was present in frozen tissue, followed by ultrasound accelerated formalin fixed tissue, then conventional formalin fixed tissue.
Biospecimens
Preservative Types
- Formalin
- Frozen
Diagnoses:
- Not specified
- Autopsy
Platform:
Analyte Technology Platform RNA RT-PCR RNA Real-time qRT-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 220 bp
489 bp
811 bp
Biospecimen Preservation Method of fixative delivery Immersion
Ultrasound-accelerated
Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
Snap frozen