Abstract
Relative abundance of tumour angiogenesis has been shown to be of clinical relevance in cancers of various locations such as the ovary. Nevertheless, several problems are encountered when quantifying tumour microvessels: (i) as many other tumour markers, vascularity pattern is often heterogeneous within the tumour mass and even within the same histological section. As a consequence, an adequate acquisition method must be developed for accurate field sampling. (ii) Manual microvessel counting is long, tedious and subject to poor reproducibility. Introduction in routine practice requires a fast, reproducible and reliable automatic image processing. In this study we present an original procedure combining a slide scanner image acquisition and a fully automatic image analysis sequence. The slide scanner offers the advantage of recording an image of the whole histological section for subsequent automatic blood vessel detection and hot spot area location. Microvessel density and surface fraction were measured for the whole section as well as within hot spots. Different immunostaining methods were tested in order to optimise the procedure. Moreover, the method proposed was submitted to a quality control procedure, with reference to interactive identification of microvessels at scanner level. This experiment showed that 93 to 97% of blood vessels were detected, according to the staining protocol used. Colour figures can be viewed on http://www.esacp.org/acp/2003/25‐2/kim.htm.
Highlights
It is well established that tumour growth and spread are highly dependent on neo-vascularization [14,32]
We propose in this paper a fully automatic method for tumour angiogenesis quantification over the whole histological section and inside automatically detected hot spots
Profiles obtained in the three channels along a diagonal line, drawn across the image show that this background is at the same level all over the image; it can be removed by a fixed threshold (Fig. 3)
Summary
It is well established that tumour growth and spread are highly dependent on neo-vascularization [14,32]. Some authors have found the stereological Chalkley point counting method, more reliable [15,23] Another step towards an objective quantification of tumour angiogenesis was proposed by introducing semiautomatic procedures [6,39]. Tumour tissue can contain necrotic areas and mucus which might be deeply stained by chromogens and could appear as large red or brown fields They were removed by thresholding an “excess blue image”, a grey scale image generated from the linear combination of the 3 components RGB [50] and computed as follows:
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