A procedure for tissue freezing and processing applicable to both intra-operative frozen section diagnosis and tissue banking in surgical pathology.

Author(s): Steu S, Baucamp M, von Dach G, Bawohl M, Dettwiler S, Storz M, Moch H, Schraml P

Publication: Virchows Arch, 2008, Vol. 452, Page 305-12

PubMed ID: 18253747 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study evaluated DNA, RNA, and protein yields and their suitability for molecular analysis among case-matched cancer specimens after cryopreservation by the following methods: (1) the specimen was wrapped in plastic, placed in a cryocassette and snap-frozen in liquid nitrogen for 10 seconds, (2) a closed cryocassette that contained the specimen embedded in OCT was immersed in isopentane pre-cooled to -80 degrees C. Eleven tumor specimens were assessed.

   Summary of Findings:

   RNA, DNA, and protein yields were equivalent among differentially preserved cryospecimens. RNA quality (as determined by 260/280 nm OD ratio) and RNA integrity (as determined by the ratio of 28S/18S rRNA and calculated RIN) also did not differ as a result of cryopreservation method. PCR-based amplification of extracted DNA and RNA produced clear bands of appropriate size for specimens preserved by either method. Western blot analysis and immunohistochemistry were successful for all specimens and was not influenced by preservation method.

   Biospecimens

   • Tissue - Kidney
   • Tissue - Lung
   • Tissue - Testis
   • Tissue - Uterus
   • Tissue - Ovary
   • Tissue - Breast
   • Tissue - Stomach

   Preservative Types

   • Frozen
   • OCT

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Germ Cell

   Platform:

   Analyte	Technology Platform
   DNA	Spectrophotometry
   DNA	PCR
   RNA	Spectrophotometry
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	RT-PCR
   Protein	Spectrophotometry
   Protein	Immunohistochemistry
   Protein	Western blot

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   PCR Specific	Targeted nucleic acid	von Hippel Lindau gene PAX-2 gene
   RT-PCR Specific	Targeted nucleic acid	beta-actin
   Immunohistochemistry Specific	Targeted peptide/protein	MIB-1
   Western blot Specific	Targeted peptide/protein	beta-actin
   Biospecimen Preservation	Type of fixation/preservation	OCT Snap frozen
   Biospecimen Preservation	Cooling or freezing method/ rate	Pre-cooled isopentane Liquid nitrogen

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2. Study Purpose

   Cellular morphology was assessed in case-matched cancer specimens after cryopreservation by the following methods: (1) carbon dioxide gas flowed directly over the specimen, (2) the specimen was wrapped in plastic, placed in a cryocassette and snap-frozen in liquid nitrogen for 10 seconds, (3) a closed cryocassette that contained the specimen embedded in OCT was immersed in isopentane pre-cooled to -80 degrees C. Eleven tumor specimens were assessed.

   Summary of Findings:

   Evidence of ice crystal formation was observed in all tissue specimens preserved by the carbon dioxide quick freeze method, but not other methods of cryopreservation. The authors note that preservation of tumor specimen morphology was slightly better with isopentane preservation compared to immersion in liquid nitrogen.

   Biospecimens

   • Tissue - Uterus
   • Tissue - Lung
   • Tissue - Testis
   • Tissue - Stomach
   • Tissue - Ovary
   • Tissue - Kidney
   • Tissue - Breast

   Preservative Types

   • Frozen
   • OCT

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Germ Cell

   Platform:

   Analyte	Technology Platform
   Morphology	H-and-E microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	OCT Snap frozen
   Biospecimen Preservation	Cooling or freezing method/ rate	Pre-cooled isopentane Liquid nitrogen Carbon dioxide gas

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