Simultaneous viewing of biopsies stained sequentially with H&E and AMACR/p63 immunostaining


Diagnosis of prostatic needle biopsies can be difficult due to benign mimics of carcinoma (e.g. atrophy) and borderline changes often termed as atypical small acinar proliferation (ASAP).

To discriminate these lesions from small focus carcinomas, immunohistochemistry (IHC) can be used to label basal cells of non-malignant glands (antibodies to p63), and carcinoma cells can be marked by alpha-methylacyl-CoA racemase (AMACR).

For a definitive diagnosis pathologists need to compare these immunostains with tissue morphology using standard hematoxylin and eosin staining (H&E). This web page is a demonstration of a new technique enabling comparison of H&E and IHC stains made sequentially on the same slide.  more

Study design: 30 H&E stained prostatic needle biopsies from 23 patients were digitized. The slides were destained and then immunostained with a cocktail of AMACR and p63 antibodies. The slides were scanned once more and the pairs of virtual slides were processed for simultaneous viewing, layered or side-by-side.  more

Publications: A complete description of the methodology, and the results of this study has been published in the February 2006 issue of The Journal of Urology

Click on a thumbnail image to view the virtual slide (in a new window)

Case 1

Case 2

Case 3

Case 4

Case 5

Case 1-5 | 6-11 | 12-16 | 17-23

AMACR: cyto-
plasmic marker upregulated in
prostate cancer

noma cells
p63: a trans-
cription factor, nuclear protein

basal cells of non-
malignant glands

Carcinoma: Cytoplasmic AMACR reactivity without nuclear p63

Prostatic intraepithelial neoplasia (PIN): AMACR-posi-
tivity in the presence of p63-posi-
tive basal cells

Layered Side-by-side

Quick instructions: The first time you open a virtual slide you will be prompted to download a small plug-in for your browser. It only has to be installed once, and can be uninstalled by standard procedures. You do not need to reboot your computer.

Currently the slides can be viewed on a PC with MS Internet Explorer or Firefox browsers.