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High-quality patient samples are required for reliable immunohistochemistry test outcomes that provide a significant benefit for patient care. Among the preanalytic variables in tissue handling, tissue thickness is thought to be easily controlled; however, whether the thickness of the tissue effects the staining intensity for antibody immunohistochemistry has not been quantitatively demonstrated. To investigate, we cut multiblock tissue sections of tonsil, liver, and kidney at 2, 4, 6, and 8 μm thicknesses. Interferometry measurements of the sectioned paraffin showed a

Quality and consistency in immunohistochemistry (IHC) protocol performance is aided by the use of proper control tissue with well-defined cellular elements/structures that reflect changes in protocol performance. This approach was integrated into the development of the new Dako FLEX Ready-to-Use antibodies. For each antibody, control tissue was chosen on the basis of the following three parameters: easily accessible tissue; stable antigen expression between samples of the same tissue type; and changes in protocol performance reflected by changes in staining intensity in defined cellular elements/structures within the tissue. The defined cellular elements/structures were divided into high-expression (HE) and low-expression (LE) structures. HE structures were generally stable in various protocol settings and served to monitor maximum staining capacity without background staining, whereas LE structures reflected minor changes in protocol performance. For the vast majority of antibodies, it was possib...

BACKGROUND. The aim of this study was to examine p16INK4a protein expression in ThinPrep (Cytyc Corporation, Marlborough, Mass) cervical specimens by using the CINtec p16INK4a Cytology Kit (Dako, Glostrup, Denmark). The ability of this assay to accurately identify underlying high-grade lesions was assessed by using follow-up biopsies and comparing these results with Hybrid Capture 2 (Digene, Gaithersburg, Md) high-risk HPV (hc2) results. METHODS. Three hundred ninety-eight residual ThinPrep samples were collected, and histological follow-up data were retrieved for abnormal cytology specimens. After preparation of a Papanicolaou-stained slide, a second slide was processed in preparation for p16INK4a immunostaining. High-risk human papillomavirus testing (hc2) was also performed. RESULTS. Of the 163 cytologically abnormal samples, 6-month biopsy follow-up data were available for 45% of the specimens. At initial blinded evaluation, 21 of the 26 cases with cervical intraepithelial neoplasia (CIN) II/III follow-up were positive for p16INK4a, yielding an overall diagnostic sensitivity of 81%; 29 of the 47 cases diagnosed as CIN I or less were p16INK4a negative, yielding a diagnostic specificity of 62%. In comparison, the hc2 test results indicated a diagnostic sensitivity of 100% with a diagnostic specificity of 15%. After review of selected cases with CIN II/III follow-up, 25 of 26 slides were deemed to be positive for p16INK4a, increasing the diagnostic sensitivity to 96%. CONCLUSIONS. The CINtec p16INK4a Cytology Kit, in combination with ThinPrep cervical samples, allowed clear evaluation of p16INK4a protein overexpression. Diagnostic specificity of the CINtec p16INK4a assay was significantly improved relative to hc2. To increase p16INK4a immunostaining in abnormal cells, a modified kit version with improved staining performance has been developed and is currently being evaluated. Cancer (Cancer Cytopathol) 2007. © 2007 American Cancer Society.

Dako has been an industry leader developing automated staining solutions for decades. We have invited Dako to offer perspectives and insights on the past, present, and future of automated staining systems. Experts from Dako answer questions regarding what automation can and cannot provide, how automation can improve workflow, how to secure high staining quality, gains from connection to laboratory information systems, and the application of digital pathology, LEAN and Six Sigma principles in the pathology laboratory.