Continuous-specimen-flow, high-throughput, 1-hour tissue processing. A system for rapid diagnostic tissue preparation.
Author(s): Morales AR, Essenfeld H, Essenfeld E, Duboue MC, Vincek V, Nadji M
Publication: Arch Pathol Lab Med, 2002, Vol. 126, Page 583-90
PubMed ID: 11958665 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 a rapid tissue processing method using microwave and convective heating on morphological preservation and IHC staining of fixed and unfixed specimens including uterus, breast, placenta, and small intestine among others.
Summary of Findings:
CTPM employed microwave and convective heating, solution I (isopropyl alcohol, acetone, polyethylene glycol, glacial acetic acid, dimethyl sulfoxide) and solution II (solution I plus low-viscosity mineral oil) for dehydration and clearing steps, and no additional formalin fixation. CTPM resulted in H&E and mucicarmine staining and ER immunostaining comparable to that seen in sections prepared by CPM, which included an initial 4 hour formalin fixation step. This was true for both formalin-fixed and fresh tissues. The authors note that H&E staining was slightly brighter in tissue processed using CTPM than CPM so exposure times were reduced for tissue processed with CTPM. Tissue blocks prepared by CTPM were softer than those prepared by CPM and resulted in sections that were easier to cut.
Biospecimens
Preservative Types
- Formalin
- None (Fresh)
Diagnoses:
- Not specified
- Normal
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Morphology H-and-E microscopy Morphology Macroscopic observation Morphology Light microscopy Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
None (fresh)
Biospecimen Preservation Duration of tissue/ specimen processing 65 min
11.5 h
Biospecimen Preservation Dehydration reagent Graded alcohols
Solution I and solution II
Biospecimen Preservation Clearing agent Solution I and solution II
Xylene
Biospecimen Preservation Clearing duration/condition 40 degrees C (convective), under pressure/vacuum
62 degrees C (microwave), no vacuum
Biospecimen Preservation Dehydration duration/condition 40 degrees C (convective), under pressure/vacuum
62 degrees C (microwave), no vacuum
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Study Purpose
The purpose of this study was to determine the effects of a rapid tissue processing method using microwave and convective heating on the preservation of RNA in testis, ovary, uterus, prostate gland, and breast specimens.
Summary of Findings:
RNA extracted from specimens processed by CTPM showed improved integrity with the addition of DEPC to the first three processing steps. RT-PCR amplification of a 500 bp amplicon of GAPDH was successful using RNA extracted from all 6 specimens processed by CTPM but only 3 of 6 specimens when RNA was extracted after CPM. Further, those 3 successful attempts showed less product compared with amplification using RNA extracted after CTPM.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Neoplastic - Benign
- Neoplastic - Carcinoma
- Normal
Platform:
Analyte Technology Platform RNA RT-PCR RNA Electrophoresis Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
None (fresh)
Biospecimen Preservation Duration of tissue/ specimen processing 65 min (CTPM)
11.5 h (CPM)
Biospecimen Preservation Dehydration reagent Graded alcohols
Solution I and solution II
Solution I and solution II plus DEPC
Biospecimen Preservation Clearing agent Solution I and solution II
Xylene
Solution I and solution II plus DEPC
Biospecimen Preservation Clearing duration/condition 40 degrees C (convective), under pressure/vacuum
62 degrees C (microwave), no vacuum
Biospecimen Preservation Dehydration duration/condition 40 degrees C (convective), under pressure/vacuum
62 degrees C (microwave), no vacuum