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BlueFuse Overview

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Once the microarray hybridization protocol has been carried out and the slides have been scanned, the next step is to quantify the images. Scanning produces two .tif image files. One represents the test readings and one the control readings. Both of these image files are loaded together into BlueFuse for quantification of each spot.

BlueFuse has been developed by a Cambridge based company called BlueGnome. It uses Bayesian algorithms to generate intensity values for each spot on the array. Briefly, the Cy5 and Cy3 files are loaded into the software and are overlaid onto one another. BlueFuse uses a .Gal file for each batch of arrays. This .Gal file contains array information for each gene and its position on the array. The software also allows positive and negative controls to be recorded. Once these have been set the software is ready to run.

BlueFuse also offers users to 'fuse' their quantified data. This combines spots from the same array on the basis of the estimated confidence in each ratio and the level of agreement between replicate spots. Once optimization is complete, quantification is automatically carried out via the software. The results are saved as .txt files and the files can now be loaded into an analysis analysis.

Recent advances in BlueFuse include the ability to carry out normalization and additional downstream analysis processes. With its ease of use and lack of user intervention, BlueFuse offers a good overall package for image quantification.

To view a basic protocol for BlueFuse, click here.

To view frequently asked questions on BlueFuse, click here.

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