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An Efficient Technique Based on Deep Learning for Automatic Focusing in Microscopic System

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2nd International Congress of Electrical and Computer Engineering (ICECENG 2023)

Abstract

During microscopic examination process, experts firstly determine the most appropriate focusing point by precisely moving the microscope stage in the Z axis. This manual focusing process may cause variability in results depending on hand-eye coordination and individual experience. Therefore, the development of automatic focusing offers superior quality and consistent imaging. Many researchers have studied automatic focusing systems and proposed many focus functions to extract focus information from multi-focus images. Nevertheless, these focusing systems still consist of several significant limitations such as requiring a supplementary material, longer running time, producing different performance depending on the sample and magnification objective. In this study, an efficient technique based on deep learning is proposed to automatically determine the focused image during microscopic examination processes to minimize these limitations in the literature. This technique takes a sequence of images with the same field of view and different focuses as input and gives the image with maximum focusing as output. The proposed technique is compared with other automatic focusing techniques on the literature. In this study, novel multi-focus image sequences obtained from liver, intestine, heart, kidney, and lung samples is prepared to evaluate the performance of automatic focusing techniques. These sequences are obtained by scanning with 10× and 40× magnification objectives. Running time, accuracy, number of local maximum points, range, and noise levels are used as evaluation criteria in this study. According to evaluation criteria results obtained in this study, it has been proven that the proposed technique provides better performance than other automatic focusing techniques.

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References

  1. Dogan, H., Ekinci, M.: Automatic panorama with auto-focusing based on image fusion for microscopic imaging system. Signal Image Video Process. 8, 5–20 (2014)

    Article  Google Scholar 

  2. Wang, C., Huang, Q., Cheng, M., Ma, Z., Brady, D.J.: Intelligent autofocus. arXiv preprint arXiv:2002 12389 (2020)

    Google Scholar 

  3. Doğan, H., Baykal, E., Ekinci, M., Ercin, M.E., Ersöz, Ş.: Determination of optimum auto focusing function for cytopathological assessment processes. In: Medical Technologies National Congress (TIPTEKNO), pp. 1–4. IEEE, Trabzon (2017)

    Google Scholar 

  4. Shi, H., Shi, Y., Li, X.: Study on auto-focus methods of optical microscope. In: 2nd Int. Conf. on Circuits, System and Simulation (ICCSS 2012), vol. 46. IPCSIT (2012)

    Google Scholar 

  5. Santos, A., Ortiz De Solorzano, C., Vaquero, J.J., Pena, J.M., Malpica, N., del Pozo, F.: Evaluation of autofocus functions in molecular cytogenetic analysis. J. Microsc. 188(3), 264–272 (1997)

    Article  Google Scholar 

  6. Rudnaya, M.E., Mattheij, R.M.M., Maubach, J.M.L.: Evaluating sharpness functions for automated scanning electron microscopy. J. Microsc. 240(1), 38–49 (2010)

    Article  MathSciNet  Google Scholar 

  7. Saini, G., Panicker, R.O., Soman, B., Rajan, J.: A comparative study of different auto-focus methods for mycobacterium tuberculosis detection from brightfield microscopic images. In: IEEE Distributed Computing, VLSI, Electrical Circuits and Robotics (DISCOVER), pp. 95–100, Mangalore (2016)

    Google Scholar 

  8. Pertuz, S., Puig, D., Garcia, M.A.: Analysis of focus measure operators for shape-from-focus. Pattern Recogn. 46(5), 1415–1432 (2013)

    Article  Google Scholar 

  9. Geusebroek, J.M., Cornelissen, F., Smeulders, A.W., Geerts, H.: Robust autofocusing in microscopy. Cytom.: J. Int. Soc. Anal. Cytol. 39, 1–9 (2000)

    Article  Google Scholar 

  10. Ahmad, M.B., Choi, T.S.: Application of three-dimensional shape from image focus in LCD/TFT displays manufacturing. IEEE Trans. Consum. Electron. 53(1), 1–4 (2007)

    Article  Google Scholar 

  11. Huang, W., Jing, Z.: Evaluation of focus measures in multi-focus image fusion. Pattern Recogn. Lett. 28(4), 493–500 (2007)

    Article  Google Scholar 

  12. Nayar, S.K.: Shape from focus system for rough surfaces. In: Physics-Based Vision: Principles and Practice: Radiometry, vol. 1, pp. 347–360. CRC Press, New York (1993)

    Google Scholar 

  13. Pech-Pacheco, J.L., Cristóbal, G., Chamorro-Martinez, J., Fernández-Valdivia, J.: Diatom autofocusing in brightfield microscopy: a comparative study. In: Proceedings 15th International Conference on Pattern Recognition. ICPR-2000, vol. 3, pp. 314–317. IEEE, Barcelona (2000)

    Chapter  Google Scholar 

  14. Thelen, A., Frey, S., Hirsch, S., Hering, P.: Improvements in shape-from-focus for holographic reconstructions with regard to focus operators, neighborhood-size, and height value interpolation. IEEE Trans. Image Process. 18(1), 151–157 (2008)

    Article  MathSciNet  Google Scholar 

  15. An, Y., Kang, G., Kim, I.J., Chung, H.S., Park, J.: Shape from focus through Laplacian using 3D window. In: In 2008 Second International Conference on Future Generation Communication and Networking, vol. 2, pp. 46–50. IEEE, Hainan (2008)

    Google Scholar 

  16. Yan, T., Hu, Z., Qian, Y., Qiao, Z., Zhang, L.: 3D shape reconstruction from multifocus image fusion using a multidirectional modified Laplacian operator. Pattern Recogn. 98, 107065 (2020)

    Article  Google Scholar 

  17. Yap, P.T., Raveendran, P.: Image focus measure based on Chebyshev moments. IEE Proc.-Vision, Image Signal Process. 151(2), 128–136 (2004)

    Article  Google Scholar 

  18. Wee, C.Y., Paramesran, R.: Measure of image sharpness using eigenvalues. Inf. Sci. 177(12), 2533–2552 (2007)

    Article  Google Scholar 

  19. Lee, S.Y., Kumar, Y., Cho, J.M., Lee, S.W., Kim, S.W.: Enhanced autofocus algorithm using robust focus measure and fuzzy reasoning. IEEE Trans. Circuits Syst. Video Technol. 18(9), 1237–1246 (2008)

    Article  Google Scholar 

  20. Lee, S.Y., Yoo, J.T., Kumar, Y., Kim, S.W.: Reduced energy-ratio measure for robust autofocusing in digital camera. IEEE Signal Process. Lett. 16(2), 133–136 (2009)

    Article  Google Scholar 

  21. Shen, C.H., Chen, H.H.: Robust focus measure for low-contrast images. In: 2006 Digest of Technical Papers International Conference on Consumer Electronics, pp. 69–70. IEEE, Las Vegas (2006)

    Chapter  Google Scholar 

  22. Ali, U., Mahmood, M.T.: 3D shape recovery by aggregating 3D wavelet transform-based image focus volumes through 3D weighted least squares. J. Math. Imaging Vis. 62, 54–72 (2020)

    Article  MathSciNet  Google Scholar 

  23. Xie, H., Rong, W., Sun, L.: Construction and evaluation of a wavelet-based focus measure for microscopy imaging. Microsc. Res. Tech. 70(11), 987–995 (2007)

    Article  Google Scholar 

  24. Shirvaikar, M.V.: An optimal measure for camera focus and exposure. In: Thirty-Sixth Southeastern Symposium on System Theory, pp. 472–475. IEEE, Atlanta (2004)

    Google Scholar 

  25. Helmli, F.S., Scherer, S.: Adaptive shape from focus with an error estimation in light microscopy. In: 2nd International Symposium on Image and Signal Processing and Analysis, pp. 188–193. IEEE Cat, Pula (2001)

    Google Scholar 

  26. Mahmood, F., Mahmood, J., Zeb, A., Iqbal, J.: 3D shape recovery from image focus using Gabor features. In: Tenth International Conference on Machine Vision, pp. 368–375 (2018). https://doi.org/10.1117/12.2309440

    Chapter  Google Scholar 

  27. Nanda, H., Cutler, R.: Practical calibrations for a real-time digital omnidirectional camera. CVPR Tech. Sketch. 20(2), 1–4 (2001)

    Google Scholar 

  28. Minhas, R., Mohammed, A.A., Wu, Q.J.: Shape from focus using fast discrete curvelet transform. Pattern Recogn. 44(4), 839–853 (2011)

    Article  Google Scholar 

  29. Lorenzo, J., Castrillon, M., Méndez, J., Deniz, O.: Exploring the use of local binary patterns as focus measure. In: International Conference on Computational Intelligence for Modelling Control & Automation, pp. 855–860, Vienna (2008)

    Google Scholar 

  30. Fan, T., Yu, H.: A novel shape from focus method based on 3D steerable filters for improved performance on treating textureless region. Opt. Commun. 410, 254–261 (2018)

    Article  Google Scholar 

  31. Minhas, R., Mohammed, A.A., Wu, Q.M., Sid-Ahmed, M.A.: 3D shape from focus and depth map computation using steerable filter. In: International Conference Image Analysis and Recognition, pp. 573–583. Springer, Berlin, Heidelberg (2009)

    Google Scholar 

  32. Xia, X., Yao, Y., Liang, J., Fang, S., Yang, Z., Cui, D.: Evaluation of focus measures for the autofocus of line scan cameras. Optik. 127(19), 7762–7775 (2016)

    Article  Google Scholar 

  33. Liu, Z., Lv, Q., Yang, Z., Li, Y., Lee, C.H., Shen, L.: Recent progress in transformer-based medical image analysis. Comput. Biol. Med. 164, 107268 (2023)

    Article  Google Scholar 

  34. Li, J., Chen, J., Tang, Y., Wang, C., Landman, B.A., Zhou, S.K.: Transforming medical imaging with transformers? A comparative review of key properties, current progresses, and future perspectives. Med. Image Anal. 85, 102762 (2023)

    Article  Google Scholar 

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Acknowledgments

We thank Karadeniz Technical University Drug and Pharmaceutical Technology Application & Research Center for their support. This study was supported by a grant from The Scientific and Technological Research Council of Turkiye (TUBITAK) (Project no. 1919B012203634).

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Dogu, F.T., Dogan, H., Dogan, R.O., Ay, I., Sezen, S.F. (2024). An Efficient Technique Based on Deep Learning for Automatic Focusing in Microscopic System. In: Seyman, M.N. (eds) 2nd International Congress of Electrical and Computer Engineering . ICECENG 2023. EAI/Springer Innovations in Communication and Computing. Springer, Cham. https://doi.org/10.1007/978-3-031-52760-9_17

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  • DOI: https://doi.org/10.1007/978-3-031-52760-9_17

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