Groenen P J T A, Blokx W A M, Diepenbroek C, Burgers L, Visinoni F, Wesseling P & van Krieken J H J M
(2011) Histopathology59, 1–7 Preparing pathology for personalized medicine: possibilities for improvement of the pre‐analytical phase With the introduction of new biological agents for cancer treatment enabling ‘personalized medicine’, treatment decisions based on the molecular features of the tumour are more common. Consequently, tissue evaluation in tumour pathology is becoming increasingly based on a combination of classical morphological and molecular analysis. The results of diagnostic tests rely not only on the quality of the method used but, to a large extent, also on the quality of specimens, which is dependent on the pre‐analytical procedures and storage. With the introduction of predictive immunohistochemical and molecular tests in clinical pathology, improvement and standardization of pre‐analytical procedures has become crucial. The aim of this review is to increase awareness with regard to tissue handling and for standardization of the pre‐analytical phase of a diagnostic process. In addition, several processing steps in tissue handling that need to be improved in order to obtain the quality needed for modern molecular medicine will be discussed. Optimal, standardized procedures are crucial if a high standard of test results is to be achieved, which is what each patient deserves.

Three-dimensional (3D) visualization of microscopic structures may provide useful information about the exact 3D configuration, and offers a useful tool to examine the spatial relationship between different components in tissues. A promising field for 3D investigation is the microvascular architecture in normal and pathological tissue, especially because pathological angiogenesis plays a key role in tumor growth and metastasis formation. This paper describes an improved method for 3D reconstruction of microvessels and other microscopic structures in transmitted light microscopy. Serial tissue sections were stained for the endothelial marker CD34 to highlight microvessels and corresponding images were selected and aligned. Alignment of stored images was further improved by automated non-rigid image registration, and automated segmentation of microvessels was performed. Using this technique, 3D reconstructions were produced of the vasculature of the normal brain. Also, to illustrate the complexity of tumor vasculature, 3D reconstructions of two brain tumors were performed: a hemangioblastoma and a glioblastoma multiforme. The possibility of multiple component visualization was shown in a 3D reconstruction of endothelium and pericytes of normal cerebellar cortex and a hemangioblastoma using alternate staining for CD34 and alpha-smooth muscle actin in serial sections, and of a GBM using immunohistochemical double staining. In conclusion, the described 3D reconstruction procedure provides a promising tool for simultaneous visualization of microscopic structures.

In the 1970s, Wellings developed and reported extensively on a technique for a three-dimensional (3D) analysis of breast lesions. Drawbacks of this subgross sampling technique were that it was laborious, rather time-consuming and only allowed prospective studies. Furthermore, the stereomicroscopic aspect of the lesions studied was not diagnostic and each sample had to be studied histologically after paraffin embedding to determine the diagnosis. The present study introduces an original method enabling the exploration of the 3D structure of the mammary glandular tree from a paraffin-embedded sample. This procedure is quicker than the Wellings' technique, permits retrospective study and enables a 3D analysis of previously identified histological structures. Stereomicroscopic aspects of non-malignant lesions such as single multiple or metaplastic cysts, adenosis, ductal-lobular hyperplasia and malignant in situ neoplasms are illustrated. Our results confirm Wellings' concept that most minimal lesions arise in the terminal ductulo-lobular units. We also show that ductal carcinoma in situ may grow continuously by extending through the glandular tree but may also have a multifocal or stepwise progression in some cases.

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