Extended Depth-of-Field for Color Images in Light Microscopy: Image Fusion and 3D Visualization

Bright-field microscopy suffers from a relatively small depth-of-field. Typically, the specimen’s profile covers a range larger than the depth-of-field, and parts of the specimen that lie outside the object plane appear blurred. The specimen can be ‘scanned’ by moving the object along the optical axis, and different images will contain different areas that are sharp. The purpose of image fusion is to combine those images into one single image with an extended depth-of-field. One promising method is based on the wavelet transform. We show how the wavelet-based image fusion technique can be successfully applied to obtain a sharp composite color image. The selection of the proper type of wavelets can improve the results. We also introduce a way to apply this technique for color images without introducing false colors. Finally, we show how height information on the specimen, which we obtain as a side product from our algorithm, can be used for 3D visualization.