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Asian Conference on Intelligent Information and Database Systems

ACIIDS 2015: Intelligent Information and Database Systems pp 297–306Cite as

Automated Analysis of Images from Confocal Laser Scanning Microscopy Applied to Observation of Calcium Channel Subunits in Nerve Cell Model Line Subjected Electroporation and Calcium

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Part of the book series: Lecture Notes in Computer Science ((LNAI,volume 9012))

Abstract

We assess a possibility of applying automated image analysis to immunofluorescence microphotographs from confocal laser scanning microscopy (CLSM). Several modes of automated analysis were tested to inspect differentiation of voltage dependent calcium channel subunits from a model of nerve cells PC12 (rat pheochromocytoma) subjected to electroporation (EP), with regard to extracellular calcium level. The objective of the experiments was evaluating sensitivity of the channel expression to the presence of calcium and electroporation voltage. For this purpose non-selective nanopores of a controlled conductivity were generated in the cell membrane using electroporation, at physiological or increased extracellular calcium concentrations. Introduction of Ca2+ into the cells was possible through electropores and physiological voltage-dependent calcium channels. Two subunits of calcium channel (α1H and α1G) were immunofluorescentically stained and the automated analysis of changes in cellular morphology was performed, based on comparative assessment of the fluorescence signal. The automated analysis allowed apparently higher observation capabilities. The results showed morphological changes in the channel subunits and higher expression of the channel, following its exposition to electric field or calcium.

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References

  1. Kanade, T., et al.: Cell image analysis: Algorithms, system and applications. In: 2011 IEEE Workshop on Applications of Computer Vision (WACV), IEEE (2011)

    Google Scholar 

  2. Wählby, C., Erlandsson, F., Bengtsson, E., Zetterberg, A.: Sequential immunofluorescence staining and image analysis for detection of large numbers of antigens in individual cell nuclei. Cytometry 47(1), 32–41 (2002)

    Article  Google Scholar 

  3. Yin, Z., Su, H., Ker, E., Li, M., Li, H.: Cell-Sensitive Microscopy Imaging for Cell Image Segmentation. In: Golland, P., Hata, N., Barillot, C., Hornegger, J., Howe, R. (eds.) MICCAI 2014, Part I. LNCS, vol. 8673, pp. 41–48. Springer, Heidelberg (2014)

    Chapter  Google Scholar 

  4. Liu, H., Felix, R., Gurnett, C., De Waard, M., Witcher, D.R., Campbell, K.P.: Expression and Subunit Interaction of Voltage-Dependent Ca2+ Channels in PC12 Cells. The Journal of Neuroscience 16(23), 7557–7565 (1996)

    Google Scholar 

  5. Beam, K.G., Adams, B.A., Niidome, T., Numa, S.: Tanabe T Function of a truncated dihydropyridine receptor as both voltage sensor and calcium channel. Nature 360, 169–171 (1992)

    Article  Google Scholar 

  6. Ghosh, A., Ginty, D.D., Bading, H., Greenberg, M.E.: Calcium regulation of gene expression in neuronal cells. J. Neurobiol. 25, 294–303 (1994)

    Article  Google Scholar 

  7. Dunlap, K.: Luebke JI, Turner TJ Exocytotic Ca2+ channels in mammalian central neurons. Trends. Neurosci. 18, 89–96 (1995)

    Article  Google Scholar 

  8. Schindelin, J., Arganda-Carreras, I., Frise, E., Kaynig, V., Longair, M., Pietzsch, T., Preibisch, S., Rueden, C., Saalfeld, S., Schmid, B., Tinevez, J.Y., White, D.J., Hartenstein, V., Eliceiri, K., Tomancak, P., Cardona, A.: Fiji: an open-source platform for biological-image analysis. Nature Methods 9(7), 676–682 (2012). doi:10.1038/nmeth.2019

    Article  Google Scholar 

  9. R Development Core Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0, URL (2008). http://www.R-project.org

  10. Huang, L.-K., Wang, M.-J.J.: Image thresholding by minimizing the measure of fuzziness. Pattern Recognition 28(1), 41–51 (1995)

    Article  Google Scholar 

  11. Prewitt, J.M.S., Mendelsohn, M.L.: The analysis of cell images. Annals of the New York Academy of Sciences 128, 1035–1053 (1966)

    Article  Google Scholar 

  12. Li, C.H., Lee, C.K.: Minimum Cross Entropy Thresholding. Pattern Recognition 26(4), 617–625 (1993)

    Article  Google Scholar 

  13. Kapur, J.N., Sahoo, P.K., Wong, A.C.K.: A New Method for Gray-Level Picture Thresholding Using the Entropy of the Histogram. Graphical Models and Image Processing 29(3), 273–285 (1985)

    Article  Google Scholar 

  14. Kittler, J., Illingworth, J.: Minimum error thresholding. Pattern Recognition 19 (1986)

    Google Scholar 

  15. Tsai, W.: Moment-preserving thresholding: a new approach. Computer Vision, Graphics, and Image Processing 29, 377–393 (1985)

    Article  Google Scholar 

  16. Otsu, N.: A threshold selection method from gray-level histograms. IEEE Trans. Sys., Man Cyber. 9, 62–66 (1979). doi:10.1109/TSMC.1979.4310076

    Article  Google Scholar 

  17. Doyle, W.: Operation useful for similarity-invariant pattern recognition. Journal of the Association for Computing Machinery 9, 259–267 (1962). doi:10.1145/321119.321123

    Article  MATH  Google Scholar 

  18. Shanbhag, A.G.: Utilization of information measure as a means of image thresholding. Graph. Models Image Process. (Academic Press, Inc.) 56(5), 414–419, ISSN 1049-9652 (1994)

    Google Scholar 

  19. Zack, G.W., Rogers, W.E., Latt, S.A.: Automatic measurement of sister chromatid exchange frequency. J. Histochem. Cytochem 25(7), 741–53. PMID 70454 (1977)

    Google Scholar 

  20. Yen, J.C., Chang, F.J., Chang, S.: A New Criterion for Automatic Multilevel Thresholding. IEEE Trans. on Image Processing 4(3), 370–378, ISSN 1057-7149, (1995). doi:10.1109/83.366472

  21. Ridler, T.W., Calvard, S.: Picture thresholding using an iterative selection method. IEEE Transactions on Systems, Man and Cybernetics 8, 630–632 (1978)

    Article  Google Scholar 

  22. Henriques, R., Lelek, M., Fornasiero, E.F., Valtorta, F.: Christophe Zimmer & Musa M Mhlanga. QuickPALM: 3D real-time photoactivation nanoscopy image processing in ImageJ. Nature Methods 7, 339–340. (2010). doi:10.1038/nmeth0510-339

  23. Kullback, S.: Information Theory and Statistics. Dover, New York (1968)

    Google Scholar 

  24. Puzicha, J., Hofmann, T., Buhmann, J.M.: Non-parametric similarity measures for unsupervised texture segmentation and image retrieval. In: Proceedings of CVPR (June 1997)

    Google Scholar 

  25. Rubner, Y., Tomasi, C., Guibas, L.J.: A metric for distributions with applications to image databases. In: IEEE International Conference on Computer Vision, pp. 59–66 (January 1998)

    Google Scholar 

  26. Klockner, U., Lee, J.H., Cribbs, L.L., Daud, A., Hescheler, J., Pereverzev, A., Perez-Reyes, E., Schneider, T.: Comparison of the Ca2+ currents induced by expression of three cloned α1 subunits, α1G, α1H and α1I, of low-voltage-activated T-type Ca2+ channels. Eur J Neurosci 11, 4171–4178 (1999)

    Article  Google Scholar 

  27. Tang, L.: Electroporation for drug and gene delivery in cancer therapy. Curr. Drug. Metab. 14(3), 271 (2013)

    Article  Google Scholar 

  28. Gibot, L., Wasungu, L., Teissié, J., Rols, M.P.: Antitumor drug delivery in multicellular spheroids by electropermeabilization. J. Control Release 28, 167(2), 138–47 (2013)

    Google Scholar 

  29. Kulbacka, J., Daczewska, M., Dubińska-Magiera, M., Choromańska, A., Rembiałkowska, N., Surowiak, P., Kulbacki, M., Kotulska, M., Saczko, J.: Doxorubicin delivery enhanced by electroporation to gastrointestinal adenocarcinoma cells with P-gp overexpression. Bioelectrochemistry. pii: S1567–5394(14) 00065-6 (April 4, 2014)

    Google Scholar 

  30. Kulbacka, J., Kotulska, M., Rembiałkowska, N., Choromańska, A., Kamińska, I., Garbiec, A., Rossowska, J., Daczewska, M., Jachimska, B., Saczko, J.: Cellular stress induced by photodynamic reaction with CoTPPS and MnTMPyPCl5 in combination with electroporation in human colon adenocarcinoma cell lines (LoVo and LoVoDX). Cell Stress Chaperones 18(6), 719–731 (2013)

    Article  Google Scholar 

  31. Nesin, V., Bowman, A.M., Xiao, S., Pakhomov, A.G.: Cell permeabilization and inhibition of voltage-gated Ca(2+) and Na(+) channel currents by nanosecond pulsed electric field. Bioelectromagnetics 33(5), 394–404 (2012)

    Article  Google Scholar 

  32. Pakhomov, A.G., Kolb, J., White, J., Shevin, R., Pakhomova, O.N., Schoenbach, K.S.: Membrane effects of ultrashort (nanosecond) electric stimuli. Society for Neuroscience 37th Annual Meeting; Nov. 2–7, 2007; San Diego, CA. 2007a. 2007 Neuroscience Meeting Planner CD-ROM, Presentation Number: 317.14

    Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Medical Biochemistry, Medical University, Chałubińskiego 10, 50-367, Wroclaw, Poland

    Julita Kulbacka, Anna Choromańska & Jolanta Saczko

  2. Polish-Japanese Academy of Information Technology, Koszykowa 86, 02-008, Warsaw, Poland

    Marek Kulbacki & Jakub Segen

  3. Department of Animal Developmental Biology, Institute of Experimental Biology, University of Wroclaw, Wroclaw, Poland

    Magda Dubińska-Magiera

  4. Department of Biomedical Engineering, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370, Wroclaw, Poland

    Małgorzata Kotulska

Authors
  1. Julita Kulbacka
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  2. Marek Kulbacki
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  3. Jakub Segen
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  4. Anna Choromańska
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  5. Jolanta Saczko
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  6. Magda Dubińska-Magiera
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  7. Małgorzata Kotulska
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Correspondence to Julita Kulbacka .

Editor information

Editors and Affiliations

  1. Ton Duc Thang University, Ho Chi Minh city, Vietnam

    Ngoc Thanh Nguyen

  2. Wroclaw University of Technology, Wroclaw, Poland

    Bogdan Trawiński

  3. Bina Nusantara University, Jakarta, Indonesia

    Raymond Kosala

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Kulbacka, J. et al. (2015). Automated Analysis of Images from Confocal Laser Scanning Microscopy Applied to Observation of Calcium Channel Subunits in Nerve Cell Model Line Subjected Electroporation and Calcium. In: Nguyen, N., Trawiński, B., Kosala, R. (eds) Intelligent Information and Database Systems. ACIIDS 2015. Lecture Notes in Computer Science(), vol 9012. Springer, Cham. https://doi.org/10.1007/978-3-319-15705-4_29

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  • DOI: https://doi.org/10.1007/978-3-319-15705-4_29

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  • Publisher Name: Springer, Cham

  • Print ISBN: 978-3-319-15704-7

  • Online ISBN: 978-3-319-15705-4

  • eBook Packages: Computer ScienceComputer Science (R0)

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