Immunohistochemical detection of progesterone receptors in archival breast cancer.

Author(s): Mote PA, Leary JA, Clarke CL

Publication: Biotech Histochem, 1998, Vol. 73, Page 117-27

PubMed ID: 9674881 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 slide type, adhesive, and section thickness on section retention of breast carcinomas and antigen retrieval techniques on PR IHC in breast carcinomas and normal myometrium.

   Summary of Findings:

   Heat-induced antigen retrieval was necessary for PR immunoreactivity, and autoclaving for 20 minutes was superior to microwave pretreatment for an equivalent amount of time (p=0.009). The hPRa 7 antibody was able to detect PR after autoclave pretreatment for 5-10 minutes and staining intensity increased after 30 minutes of antigen retrieval. However, the hPRa 6 antibody yielded only weak immunostaining after antigen retrieval of less than 20 minutes. Section loss (absent, damaged, or frayed) occurred more often with section thicknesses of 4 uM compared with 2 uM. After antigen retrieval by autoclaving for 20 minutes, significantly more sections were retained on Superfrost Plus slides than Histogrip or Silane coated slides (p=0.0001), the latter two of which performed similarly. The use of additional adhesives did not improve section retention for Histogrip slides while Para Pen and Mayer albumin each improved section retention for Silane and Superfrost Plus slides. Mayer albumin was the best adhesive, and the best combination of slide and adhesive was Superfrost Plus slides with Mayer albumin.

   Biospecimens

   • Tissue - Breast
   • Tissue - Uterus

   Preservative Types

   • Formalin

   Diagnoses:

   • Normal
   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry
   Morphology	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Aliquot size/volume	2 uM thick section 4 uM thick section
   Biospecimen Aliquots and Components	Type of slide	Histogrip Silane Superfrost Plus
   Biospecimen Aliquots and Components	Tissue section adhesion	Uncoated Mayer albumin adhesive Para Pen
   Analyte Extraction and Purification	Temperature of heat-induced retrieval	Autoclave Microwave
   Analyte Extraction and Purification	Antigen retrieval	0 min 5 min 10 min 15 min 20 min 30 min 36 min
   Immunohistochemistry Specific	Type of antibody	hPRa 7 hPRa 6

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