Abstract
Non-invasive and marker-free monitoring of living cells can be accomplished by vector contrast scanning acoustic microscopy. In this paper, the signal processing required for creating time-lapse movies of mesenchymal stem cells is discussed. This includes electronic signal processing, autofocusing and image processing. Prior to each recorded image the focusing transducer is moved away from the sample until no echo signal is received. This allows direct measurement of the offset vector caused by internal lens echoes. The offset vector can then be subtracted from the following vector-contrast image. For subsequent autofocusing the transducer is moved closer to the sample until the maximum of the signal in reflection is passed. The transducer position for the maximum reflected signal is determined by respective software and adjusted accordingly. Autofocusing is a requirement for tiled scans where a piezo-scanner and an automatic microscope stage are combined to increase the field of view. As there are typically thousands of images involved in a single movie, batch image processing routines are required. Customized plugins for ImageJ were developed to combine specialized functions for vector contrast data processing with standard image processing capabilities. The motility of a population of ovine mesenchymal stem cells was continuously recorded for 8 h. The detection scheme including experimental details is presented and applications including time-lapse imaging are demonstrated and discussed.
Keywords
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Hildebrand, J.A., Ruger, D., Johnston, R.N., Quate, C.F.: Acoustic microscopy of living cells. PNAS 78(3), 1656–1660 (1981)
Cross, S., Jin, Y., Rao, J., Gimzewski, J.: Nanomechanical analysis of cells from cancer patients. Nat. Nanotechnol. 2, 780–783 (2007)
Grill, W., Hillman, K., Würz, K.U., Wesner, J. In: Briggs, A., Arnold, W. (ed.) Advances in Acoustic Microscopy, vol. 2, pp. 167–218. Plenum Press, New York (1996)
Briggs, G.A.D., Wang, J., Gundle, R.: Quantitative acoustic microscopy of individual living human cells. J. Microsc. 172, 3–12 (1993)
A-Hassan, E., Heinz, W.F., Antonik, M.D., D’Costa, N.P., Nageswaran, S., Schoenenberger, C-A., Hoh, J.H.: Relative microelastic mapping of living cells by atomic force microscopy. Biophys. J. 74, 1564–1578 (1998)
Fabry, B., Maksym, G.N., Butler, J.P., Glogauer, M., Navajas, D., Taback, N.A., Millet, E.J., Fredberg, J.J.: Time scale and other invariants of integrative mechanical behavior in living cells. Phys. Rev. E 68(4), 041914 (2003)
Wottawah, F., Schinkinger, S., Lincoln, B., Ananthakrishnan, R., Romeyke, M., Guck, J., Käs, J.: Optical rheology of biological cells. Phys. Rev. Lett. 94(098103), 1–4 (2005)
Kundu, T., Bereiter-Hahn, J., Karl, I.: Cell property determination from the acoustic microscope generated voltage versus frequency curves. Biophys. J. 78, 2270–2279 (2000)
Kundu, T., Bereiter-Hahn, J., Hillmann, K.: Measuring elastic properties of cells by evaluation of scanning acoustic microscopy v(z) values using simplex algorithm. Biophys. J. 59, 1194–1207 (1991)
Bereiter-Hahn, J., Blase, C.: Ultrasonic characterization of biological cells. In: Kundu, T. (ed.) Ultrasonic Nondestructive evaluation, pp. 725–759. CRC Press, Boca Raton, FL (2003)
Liang, K.K., Bennett, S.D., Kino, G.S.: , Precision phase measurement with short tone burst signals in acoustic microscopy. Rev. Sci. Instrum. 57, 446–452 (1986)
Lemons, R.A., Quate, C.F.: Acoustic microscope - scanning version. Appl. Phys. Lett. 24(4), 163–165 (1974)
Grill, W., Hillmann, K., Kim, T.J., Lenkeit, O., Ndop, J., Schubert, M.: Scanning acoustic microscopy with vector contrast. Physica B 263, 553–558 (1999)
Lemor, R.M., Weiss, E.C., Pilarczyk, G., Zinin, P.V.: Measurements of elastic properties of cells using high-frequency time-resolved acoustic microscopy. IEEE Ultrasonics Symposium, pp. 762–765 (2003)
Kamanyi, A., Ngwa, W., Betz, T., Wannemacher, R., Grill, W.: Combined phase-sensitive acoustic microscopy and confocal laser scanning microscopy. Ultrasonics 44, e1295–e1300 (2006)
Schenkl, S., Weiss, E.C., Stracke, F., Sauer, D., Stark, M., Riemann, I., Lemor, R.M., König, K.: In-vivo observation of cells with a combined high-resolution multiphoton-acoustic scanning microscope. Microsc. Res. Tech. 70, 476–480 (2007)
von Buttlar, M., Twerdowski, E., Grill, W.: Offset correction for scanning acoustic microscopy with phase contrast. Proc. Int. Congr. Ultras., Paper ID 1698 (2007)
Rasband, W.S.: ImageJ, U. S. National Institute of Health, Bethesda, Maryland, USA, http://rsb.info.nih.gov/ij/ (1997–2009)
Zscharnack, M., Poesel, C., Galle, J., Bader, A.: Low oxygen expansion improves subsequent chondrogenesis of ovine bone-marrow-derived mesenchymal stem cells in collagen type I hydrogel. Cells Tissues Organs. doi:10.1159/000178024 (2008)
Acknowledgments
We would like to thank Matthias Zscharnak, Claudia Pösel and Frank Peinemann for providing the cells and the Federal Ministry of Education and Research (BMBF grant 0313836, MS CartPro) for financial support.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media B.V.
About this paper
Cite this paper
von Buttlar, M., Mohamed, E., Grill, W. (2011). Signal Processing for Time-Lapse Cell Imaging with Vector-Contrast Scanning Acoustic Microscopy. In: André, M., Jones, J., Lee, H. (eds) Acoustical Imaging. Acoustical Imaging, vol 30. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-3255-3_17
Download citation
DOI: https://doi.org/10.1007/978-90-481-3255-3_17
Published:
Publisher Name: Springer, Dordrecht
Print ISBN: 978-90-481-3254-6
Online ISBN: 978-90-481-3255-3
eBook Packages: Physics and AstronomyPhysics and Astronomy (R0)