Theory and implementation of an electronic, automated measurement system for images obtained from immunohistochemically stained slides

Objective: To develop and implement an Internet-based, automated image measurement system for immunohistochemically stained slides including fluorescence images in online and off-line modes.

Study Design: An image analyzing system was developed that automatically measures digitized images obtained from immunohistochemically stained slides. It is divided into a common server platform and a specific image quantification system based upon DIAS (University of Jena). After registration, the user fills in an input data form and attaches images to be measured. The server periodically transfers the data to the measurement system. The measurement works on dynamic thresholding and active sampling of objects visualized by fluorescence and conventional chromogens. It includes stereologic algorithms, object quantification, syntactic structure analysis and quality assurance.

Results: The system has been tested for diaminobenzidene, alkaline phosphatase and fluorescence images (FITC, etc.). The reproducibility and stability of the system are > 98%. The series of successfully measured images comprises > 1,000 images in total in the online and off-line modes.

Conclusion: An Internet-based automated image measurement system has been developed that offers worldwide access to the major requests for quantification of immunohistochemically stained slides-tissue array analysis, nuclear stains (MIB, hormones), membrane stains (CerbB2), vascularization and fluorescence in situ hybridization.