An improved processing method for breast whole-mount serial sections for three-dimensional histopathology imaging.

Author(s): Sun L, Wang D, Zubovits JT, Yaffe MJ, Clarke GM

Publication: Am J Clin Pathol, 2009, Vol. 131, Page 383-92

PubMed ID: 19228644 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of microwave assisted processing and shortened conventional processing schedules on the preservation of morphology and quality of immunostaining in breast tissue. Pathology inked specimens were suspended in agar, chilled on ice for 1-2 h, and sliced into 4 mm thick slices prior to fixation.

   Summary of Findings:

   Serial section yields, calculated as the percentage of sections free from sectioning artifacts from a contiguous series of 10, were 100% for section area sizes of 4000 and 6000 mM^2 when microwave-based tissue processing was employed. Conventional tissue processing schedules lasting 57 h and 35 h resulted in acceptable serial section yields of 70-100%, while a conventional processing schedule lasting 22 h resulted in no serial section yield for either section area size. Morphological preservation and IHC staining were equivalent for microwave-based processing and conventional tissue processing schedules. Microwave-based processing led to an average of 4% tissue shrinkage after clearing and 8% tissue shrinkage after embedding when compared to prior to fixation.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Benign
   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry
   Morphology	Macroscopic observation
   Morphology	H-and-E microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Embedding medium	Agar Paraffin
   Biospecimen Preservation	Time in fixative	16 h (room temperature) plus 20 min (microwave) 1-3 d (room temperature)
   Biospecimen Preservation	Duration of tissue/ specimen processing	22 h (microwave) 22 h (conventional) 35 h (conventional) 57 h (conventional)
   Biospecimen Preservation	Dehydration reagent	Acetone Ethanol Isopropanol
   Biospecimen Preservation	Dehydration duration/condition	4 h 50 min 14 h 22 h 36 h
   Biospecimen Preservation	Clearing duration/condition	1.5 h 2 h 4 h 6 h
   Biospecimen Preservation	Embedding duration/condition	14 h plus 40 min (microwave) plus 2 h (conventional processor) 6 h (conventional processor) 12 h (conventional processor) 15 h (conventional processor)
   Immunohistochemistry Specific	Targeted peptide/protein	ER PR Myosin-smooth muscle E-cadherin HER2/neu
   Biospecimen Aliquots and Components	Aliquot size/volume	4000 mM^2 6000 mM^2

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