Localization and Conversion of Single Cell Positions from Static High-Resolution Digital Images to Lasermicrodissector Coordinate System through Utilization of References and 2D Transformation Techniques

Abstract

laser microdissection technique for tissue excision has been developed to replace the slow, manual positioning of the laser with a roller. The method consists of the 3DHistech- P1000 digital tissue scanner, MorphCheck evaluation software, and integration of the MMI CellCut laser microdissector. To create a seamless integration between the workflows, a digital reference selection; rotational, translational, and conversion transformation of Region of Interest (ROI) coordinates from pixel-based images to the mm-based coordinate system of the laser unit; a fused database loaded with parameters (image, shape, position, unique identifier) of the analyzed slides and the selected ROIs, and XML-based communication have been developed. The software implementation has been tested on the microdissector device. The application of the method results in an automated workflow that allows laser excision with an accuracy of 2 μm and a standard deviation of 2.2 μm, which requires the selection of at least three cell clusters of ROIs to perform laser excision without live image based on a predefined static image.