Aims: To establish a three dimensional reconstruction of an invasive breast carcinoma using basic laboratory equipment to evaluate and characterise the spatial arrangement of the parenchymal cells of the breast.
Methods: One hundred and twenty eight sequential 4 μm sections (20 μm apart) of the tumour were stained immunohistochemically with an epithelial specific marker (AE1/AE3) or tumour specific marker (c-erbB-2) to reconstruct two different three dimensional images of the normal and malignant parenchymal cells. Sections were digitally imaged using a microscope, scanner, and digital camera linked to a conventional personal computer. Accurate alignment of the digitalised images was carried out using a semiautomatic graphical method of manual interaction, using the cross correlation coefficient as a goodness of fit measure, and an automatic search algorithm using the Fibonacci search algorithm for automatic alignment. The volume was reconstructed using maximum, minimum point projection and “back to front” opacity blending.
Results: The quality of the reconstructed images was distinct and perfect, providing a comprehensive and explicit view of the normal and malignant parenchymal tissues of the breast that is not possible by viewing two dimensional histological sections. Specifically, this approach showed the spatial arrangement of the tumour cells and their relation to the surrounding tissues at a high resolution.
Conclusion: This simple and reproducible approach enables the spread and infiltration of invasive carcinoma to be understood and could also be used to analyse the spatial relation between atypical hyperplastic and malignant in situ lesions of the breast.
- CLSM, confocal laser scanning microscopy
- DCIS, ductal carcinoma in situ
- NSS, normal swine serum
- TBS, Tris buffered saline
- breast cancer
- computer assisted three dimensional reconstruction
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Competing interests: Professor I O Ellis is the Medical Director of Medical Solutions PLC and has responsibility for the strategic direction and medical supervision of the company’s diagnostic pathology and laboratory services. There are no other competing interests.