Long-term preservation of antigenicity on tissue microarrays.
Tissue microarrays have facilitated the evaluation of large cohort studies; however, there is little data on the best method for preserving sections once they are cut. We assessed three methods of storing precut breast cancer microarray slides: paraffin coating and storage in a nitrogen desiccator, either alone or in combination. We tested the durability of three antigens, cytokeratin, estrogen receptor, and Ki-67 on microarrays stored under these conditions for 3 months at room temperature. Staining was assessed with both manual scoring using traditional brown stain (0-3+) as well as automated scoring using fluorescently stained sections. Staining intensity was compared to that obtained from freshly cut slides. Slides stored under ambient conditions (room temperature and air) for 3 months exhibited marked degradation of all target antigens, in some cases resulting in slides that were virtually unreadable. We found that combined paraffin coating and nitrogen storage resulted in the best preservation of antigenicity, with retention of 72-99% of the antigenicity of a freshly cut slide, depending upon the marker and detection system used. The use of either paraffin coating or nitrogen storage alone protected slides to a lesser degree.
DiVito
Kyle A
KA
Department of Pathology, Yale University, New Haven, CT, USA.
Charette
Lori A
LA
Rimm
David L
DL
Camp
Robert L
RL
D000951:Antigens, Neoplasm / Q000032:analysis*; D001943:Breast Neoplasms / Q000737:chemistry* / Q000276:immunology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis / Q000276:immunology; D019394:Ki-67 Antigen / Q000032:analysis; D009584:Nitrogen; D010232:Paraffin; D040081:Protein Array Analysis / Q000379:methods*; D011960:Receptors, Estrogen / Q000032:analysis / Q000276:immunology; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods
D000951:Antigens, Neoplasm; D019394:Ki-67 Antigen; D011960:Receptors, Estrogen; D007633:Keratins; D010232:Paraffin; D009584:Nitrogen