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Extended Depth-of-Focus for Multi-Channel Microscopy Images: A Complex Wavelet Approach

B. Forster, D. Van De Ville, J. Berent, D. Sage, M. Unser

Proceedings of the Second IEEE International Symposium on Biomedical Imaging: From Nano to Macro (ISBI'04), Arlington VA, USA, April 15-18, 2004, pp. 660-663.



Microscopy imaging often suffers from limited depth-of-focus. However, the specimen can be “optically sectioned” by moving the object along the optical axis; different areas appear in focus in different images. Extended depth-of-focus is a fusion algorithm that combines those images into one single sharp composite. One promising method is based on the wavelet transform. In this paper, we show how the wavelet-based image fusion technique can be improved and easily extended to multi-channel data. First, we propose the use of complex-valued wavelet bases, which seem to outperform traditional real-valued wavelet transforms. Second, we introduce a way to apply this technique for multi-channel images that suppresses artifacts and does not introduce false colors, an important requirement for multi-channel fluorescence microscopy imaging. We evaluate our method on simulated image stacks and give results relevant to biological imaging.


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