Improved wavelet-based microscope autofocusing for blood smears by using segmentation

Video-based autofocus has become a viable option for microscopes due to the availability of fast microcomputers and cameras that provide high frame rates. The methods proposed plot a measure of the focus vs. the frame number, commonly referred to as focus function which results in a peak when the in focus frame is reached. Recently, generic waveletbased schemes have been proposed that offer varying degrees of performance depending on the specimens being observed. The performance of these methods can be improved if the nature of the specimen being observed is known. One such scheme for blood smears based on segmentation is presented in this paper. It exploits the fact that the primary objects of interest, the Red Blood Cells (RBC), have a smooth texture. It segments the RBCs and then applies the wavelet-based focus measure. This results in a smooth focus function which permits accurate detection of the in focus frame. The proposed scheme is evaluated using several videos taken from blood smears and the results show that segmentation step improves the wavelet-based measure.

[1]  N. Otsu A threshold selection method from gray level histograms , 1979 .

[2]  I T Young,et al.  A comparison of different focus functions for use in autofocus algorithms. , 1985, Cytometry.

[3]  K Cook,et al.  Comparison of autofocus methods for automated microscopy. , 1991, Cytometry.

[4]  Michel Barlaud,et al.  Image coding using wavelet transform , 1992, IEEE Trans. Image Process..

[5]  I. Daubechies Ten Lectures on Wavelets , 1992 .

[6]  Andrew G. Dempster,et al.  Segmentation of blood images using morphological operators , 2000, Proceedings 15th International Conference on Pattern Recognition. ICPR-2000.

[7]  Kevin Barraclough,et al.  I and i , 2001, BMJ : British Medical Journal.

[8]  A. Dempster,et al.  Modification on distance transform to avoid over-segmentation and under-segmentation , 2002, International Symposium on VIPromCom Video/Image Processing and Multimedia Communications.

[9]  Georges Flandrin,et al.  Automated Detection of Working Area of Peripheral Blood Smears Using Mathematical Morphology , 2003, Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology.

[10]  Bradley J. Nelson,et al.  Wavelet-based autofocusing and unsupervised segmentation of microscopic images , 2003, Proceedings 2003 IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS 2003) (Cat. No.03CH37453).

[11]  G. A Theory for Multiresolution Signal Decomposition : The Wavelet Representation , 2004 .

[12]  Bradley J. Nelson,et al.  Autofocusing algorithm selection in computer microscopy , 2005, 2005 IEEE/RSJ International Conference on Intelligent Robots and Systems.

[13]  David M. Rubin,et al.  Automated image processing method for the diagnosis and classification of malaria on thin blood smears , 2006, Medical and Biological Engineering and Computing.

[14]  Pierre Machart Morphological Segmentation , 2009 .

[15]  Vishnu Vardhan Makkapati Segmentation based microscope autofocusing for blood smears , 2009, Medical Imaging.