Reconstruction and Elimination of Optical Microscopic Background Using Surface Fitting Method

One serious problem among the troubles to identify objects in an optical microscopic image is contour background due to non-uniform light source and various transparency of samples. To solve this problem, this paper proposed an elimination method of the contour background and compensation technique as follows. First, Otsu's optimal thresholding method extracts pixels representing background. Second, bilinear interpolation finds non-deterministic background pixels among the sampled pixels. Third, the 2D cubic fitting method composes surface function from pivoted background pixels. Fourth, reconstruction procedure makes a contour image from the surface function. Finally, elimination procedure subtracts the approximated background from the original image. To prove the effectiveness of the proposed algorithm, this algorithm is applied to the yeast Zygosaccharomyces rouxii and ammonia-oxidizing bacteria Acinetobacter sp. Labeling by this proposed method can remove some noise and is more exact than labeling by only Otsu's method. Futhermore, we show that it is more effective for the reduction of noise.