Acid Hematoxylin Is an Alternative to Feulgen Staining for Cytology Specimens and DNA Ploidy Analysis
David G Bostwick, Nikola Valkov, Rick Voyles, Michael Kulikowski, Deloar Hossain. Bostwick Laboratories, Inc., Orlando, FL
Background: Feulgen stain is the standard for nuclear image analysis and DNA ploidy analysis, but is laborious, expensive, and difficult to successfully employ with cytologic specimens owing to variable staining and loss of cells during the acid hydrolysis stage. In this report, we evaluated the utility of a potential alternative- acid hematoxylin- to Feulgen stain for cytology and DNA ploidy analysis.
Design: We undertook a split-sample comparative analysis of endometrial brush cytology samples and random void urine cytology specimens stained with Feulgen and acid hematoxylin. Brightfield digital images were analyzed for staining intensity and DNA ploidy analysis.
Results: Cell yield was invariably greater with acid hematoxylin than Feulgen stain, especially in urine samples. DNA ploidy analyses were remarkably similar, with Pearson's correlation coefficients (r) of 0.96 and 0.99 for endometrial and urine cytology specimens, respectively.
Conclusions: Acid hematoxylin had similar performance characteristics as Feulgen in DNA determination of ploidy, but was technically easier and considerably less expensive with greater cell yield for both sets of specimens.
Wednesday, March 6, 2013 9:30 AM
Poster Session V # 291, Wednesday Morning