Skip to main content
SpringerLink
Log in

Ultrasound Imaging of Apoptosis: Spectroscopic Detection of DNA-Damage Effects at High and Low Frequencies

  • Protocol
  • First Online:
DNA Damage Detection In Situ, Ex Vivo, and In Vivo

Part of the book series: Methods in Molecular Biology ((MIMB,volume 682))

Abstract

A new noninvasive method for the detection of DNA damage using mid-to high-frequency ultrasound (10–60 MHz) has been developed. Ultrasound imaging and quantitative analysis methods are used to detect cell death occurring in response to anticancer therapies in cell samples in vitro, in rat brain tissue ex vivo, and in cancer mouse models in vivo. Experimental evidence indicates that the mechanism behind this ultrasonic detection is linked to changes in the size and acoustic properties of the cell nucleus occurring with forms of cell death, and in particular apoptosis. Nuclear changes associated with cell death can result in up to 16-fold increase in ultrasound backscatter intensity and changes in spectral slope that are consistent with theoretical predictions. Furthermore, color-coded images can be generated based on specific ultrasound parameters in order to identify the regions of cell death in tumor ultrasound images with treatments. These results provide a foundation for future investigations regarding the use of ultrasound in preclinical and clinical settings to noninvasively monitor tumor responses to specific interventions that induce cell death.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Van Cruchten S., Van den Broeck W. (2002) Morphological and biochemical aspects of apoptosis, oncosis and necrosis. Anat Histol Embryol 31, 214–223.

    Article  PubMed  Google Scholar 

  2. Darzynkiewicz Z, Juan G, Li X, Gorczyca W, Murakami T, Traganos F. (1997) Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry 27, 1–20.

    Article  PubMed  CAS  Google Scholar 

  3. Darzynkiewicz Z, Juan G, Bedner E. (2001) Determining cell cycle stages by flow cytometry. Curr Protoc Cell Biol Chapter 8:Unit 8.4.

    Google Scholar 

  4. Czarnota GJ, Kolios MC, Abraham J, et al. (1999) Ultrasound imaging of apoptosis: high-resolution non-invasive monitoring of programmed cell death in vitro, in situ and in vivo. Br J Cancer 81, 520–527.

    Article  PubMed  CAS  Google Scholar 

  5. Kolios MC, Czarnota GJ, Lee M, Hunt JW, Sherar MD. (2002) Ultrasonic spectral para­meter characterization of apoptosis. Ultrasound Med Biol 28, 589–597.

    Article  PubMed  CAS  Google Scholar 

  6. Czarnota GJ, Kolios MC, Hunt JW, Sherar MD. (2002) Ultrasound imaging of apoptosis. DNA-damage effects visualized. Methods Mol Biol 203, 257–277.

    PubMed  CAS  Google Scholar 

  7. Tunis AS, Czarnota GJ, Giles A, Sherar MD, Hunt JW, Kolios MC. (2005) Monitoring structural changes in cells with high-frequency ultrasound signal statistics. Ultrasound Med Biol 31, 1041–1049.

    Article  PubMed  CAS  Google Scholar 

  8. Banihashemi B, Vlad RM, Giles A, Kolios MC, Czarnota GJ. (2008) Ultrasound imaging of apoptosis in tumour response: novel monitoring of photodynamic therapy effects. Cancer Res 68, 8590–8596.

    Article  PubMed  CAS  Google Scholar 

  9. Vlad RM, Alajez NM, Giles A, Kolios MC, Czarnota GJ. (2008) Quantitative ultrasound characterization of cancer radiotherapy effects in vitro Int J Radiat Oncol Biol Phys 72, 1236–1243.

    Article  PubMed  Google Scholar 

  10. Vlad RM, Czarnota GJ, Giles A, Sherar MD, Hunt JW, Kolios MC. (2005) High-frequency ultrasound for monitoring changes in liver tissue during preservation. Phys Med Biol 50, 197–213.

    Article  PubMed  Google Scholar 

  11. Tannock IF, Hill RP, Bristow RG, Harrington L. The basic science of oncology. New York: McGraw-Hill, 2005.

    Google Scholar 

  12. Lockshin RA, Tilly JL, Zakeri Z. When cells die: a comprehensive evaluation of apoptosis and programmed cell death. New York: Wiley-Liss, 1998.

    Google Scholar 

  13. Foster FS, Pavlin CJ, Harasiewicz KA, Christopher DA, Turnbull DH. (2000) Advances in ultrasound biomicroscopy. Ultrasound Med Biol 26, 1–27.

    Article  PubMed  CAS  Google Scholar 

  14. Foster FS, Zhang MY, Zhou YQ, et al. (2002) A new ultrasound instrument for in vivo microimaging of mice. Ultrasound Med Biol 28, 1165–1172.

    Article  PubMed  CAS  Google Scholar 

  15. Foster FS, Zhang M, Duckett AS, Cucevic V, Pavlin CJ. (2003) In vivo imaging of embryonic development in the mouse eye by ultrasound biomicroscopy. Invest Ophthalmol Vis Sci 44, 2361–2366.

    Article  PubMed  Google Scholar 

  16. Graham KC, Wirtzfeld LA, MacKenzie LT, et al. (2005) Three-dimensional high-frequency ultrasound imaging for longitudinal evaluation of liver metastases in preclinical models. Cancer Res 65, 5231–5237.

    Article  PubMed  CAS  Google Scholar 

  17. Cheung AM, Brown AS, Hastie LA, et al. (2005) Three-dimensional ultrasound biomicroscopy for xenograft growth analysis. Ultrasound Med Biol 31, 865–870.

    Article  PubMed  Google Scholar 

  18. Goertz DE, Yu JL, Kerbel RS, Burns PN, Foster FS. (2002) High-frequency Doppler ultrasound monitors the effects of antivascular therapy on tumor blood flow. Cancer Res 62, 6371–6375.

    PubMed  CAS  Google Scholar 

  19. Czarnota GJ, Kolios MC, Vaziri H, et al. (1997) Ultrasonic biomicroscopy of viable, dead and apoptotic cells. Ultrasound Med Biol 23, 961–965.

    Article  PubMed  CAS  Google Scholar 

  20. Lizzi FL, Greenebaum M, Feleppa EJ, Elbaum M, Coleman DJ. (1983) Theoretical framework for spectrum analysis in ultrasonic tissue characterization. J Acoust Soc Am 73, 1366–1373.

    Article  PubMed  CAS  Google Scholar 

  21. Lizzi FL. (1997) Ultrasonic scatterer-­property images of the eye and prostate. Proc IEEE Ultrason Symp 2, 1109–1118.

    Google Scholar 

  22. Lizzi FL, Astor M, Liu T, Deng C, Coleman DJ, Silverman RH. (1997) Ultrasonic spectrum analysis for tissue assays and therapy evaluation. Int J Imaging Syst Technol 8, 3–10.

    Article  Google Scholar 

  23. Lizzi FL, Ostromogilsky M, Feleppa EJ, Rorke MC, Yaremko MM. (1987) Relationship of ultrasonic spectral parameters to features of tissue microstructure. IEEE Trans Ultrason Ferroelectr Freq Control 34, 319–329.

    Article  PubMed  CAS  Google Scholar 

  24. Lizzi FL1, Feleppa EJ, Kaisar AS, Deng CX. (2003) Ultrasonic spectrum analysis for tissue evaluation. Pattern Recognit Lett 24, 637–658.

    Article  Google Scholar 

  25. Ursea R, Coleman DJ, Silverman RH, Lizzi FL, Daly SM, Harrison W. (1998) Correlation of high-frequency ultrasound backscatter with tumor microstructure in iris melanoma. Ophthalmology 105, 906–912.

    Article  PubMed  CAS  Google Scholar 

  26. Lizzi FL, Astor M, Feleppa EJ, Shao M, Kalisz A. (1997) Statistical framework for ultrasonic spectral parameter imaging. Ultrasound Med Biol 23, 1371–1382.

    Article  PubMed  CAS  Google Scholar 

  27. Feleppa EJ, Kalisz A, Melgar S, et al. (1996) Typing of prostate tissue by ultrasonic spectrum analysis. IEEE Trans Ultrason Ferroelectr Freq Control 43, 609–619.

    Article  Google Scholar 

  28. Lizzi FL, Astor M, Kalisz A, et al. (1996) Ultrasonic spectrum analysis for assays of different scatterer morphologies: theory and very-high frequency clinical results. Proc IEEE 1996 Ultrason Symp 2, 1155–1159.

    Article  Google Scholar 

  29. Lizzi FL, King DL, Rorke MC, et al. (1988) Comparison of theoretical scattering results and ultrasonic data from clinical liver examinations. Ultrasound Med Biol 14, 377–385.

    Article  PubMed  CAS  Google Scholar 

  30. Shung KK, Thieme GA, Ultrasonic Scattering in Biological Tissues, Insana MF, Brown DG, Chapter 4. Boca Raton, FL: CRC Press, 1993.

    Google Scholar 

  31. Insana MF, Wagner RF, Brown DG, Hall TJ. (1990) Describing small-scale structure in random media using pulse-echo ultrasound. J Acoust Soc Am 87, 179–192.

    Article  PubMed  CAS  Google Scholar 

  32. Taggart LR, Baddour RE, Giles A, Czarnota GJ, Kolios MC. (2007) Ultrasonic characterization of whole cells and isolated nuclei. Ultrasound Med Biol 33, 389–401.

    Article  PubMed  Google Scholar 

  33. Brand S, Solanki B, Foster D, Czarnota GJ, Kolios MC. (2009) Monitoring of cell death in epithelial cells using high frequency ultrasound spectroscopy. Ultrasound Med Biol 35, 482–93.

    Article  PubMed  Google Scholar 

  34. Zamble DB, Lippard SJ. (1995) Cisplatin and DNA repair in cancer chemotherapy. Trends Biochem Sci 20, 435–439.

    Article  PubMed  CAS  Google Scholar 

  35. Vlad RM, Orlova V, Hunt JW, Kolios MC, Czarnota GJ. (2008) Changes measured in the backscatter ultrasound signals during cell death can be potentially explained by an increase in cell size variance. Ultrasonic Imaging 29, 256.

    Google Scholar 

  36. Shung KK. Diagnostic ultrasound: Imaging and blood flow measurements. Boca Raton, FL: CRC Press, 2005, p. 155.

    Book  Google Scholar 

  37. Czarnota GJ, Papanicolau N, Lee J, Karshafian R, Giles A, Kolios MC. (2008) Novel low-frequency ultrasound detection of apoptosis in vitro and in vivo.. Ultrasonic Imaging 29, 237–238.

    Google Scholar 

  38. Brand S, Weiss EC, Lemor RM, Kolios MC. (2008) High frequency ultrasound tissue characterization and acoustic microscopy of intracellular changes. Ultrasound Med Biol 34, 1396–1407.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We gratefully thank Drs. Fei-Fei Liu and Deborah Foster for providing some of the cell lines used in this work. We thank Drs. Nehad Alajez, Sebastian Brand, and Anoja Giles for providing technical support. We thank Tennyson and Bradbury D. Bear for continued indefatigable scientific discourse. This research was supported by the American Institute of Ultrasound in Medicine’s Endowment for Education and Research Grant, Canadian Institutes of Health Research Strategic Training Fellowship Excellence in Radiation Research for the twenty-first century to RMV; Natural Sciences Engineering Research Council of Canada, Canada Research Chair Programme, Canadian Institutes of Health Research, Canadian Foundation of Innovation/Ontario Innovation Trust, Ryerson University to MCK; Sunnybrook Health Sciences Centre, Natural Sciences Engineering Research Council of Canada, Cancer Care Ontario Cancer Imaging Network of Ontario grants to GJC. Support from a Tier II Canada Research Chair Award and GJC from a Ontario Clinician Scientist Award from the Ontario Ministry of Health and Long-Term Care.

Author information

Authors and Affiliations

  1. Department of Medical Biophysics, Radiation Medicine Program, Princess Margaret Hospital, University of Toronto, Toronto, ON, Canada

    Roxana M. Vlad

  2. Department of Radiation Oncology and Imaging Research, Sunnybrook Health Sciences Centre, Toronto, ON, Canada

    Roxana M. Vlad & Gregory J. Czarnota

  3. Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada

    Michael C. Kolios

  4. Department of Physics, Ryerson University, Toronto, ON, Canada

    Michael C. Kolios

  5. Department of Medical Biophysics, Department of Radiation Oncology, Faculty of Medicine, University of Toronto, Toronto, ON, Canada

    Gregory J. Czarnota

Authors
  1. Roxana M. Vlad
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael C. Kolios
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gregory J. Czarnota
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gregory J. Czarnota .

Editor information

Editors and Affiliations

  1. Baylor College of Medicine, Depts. Neurosurgery and Molecular & Cell, University of Houston, Holcombe Blvd. 2002, Houston, 77030, Texas, USA

    Vladimir V. Didenko

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Vlad, R.M., Kolios, M.C., Czarnota, G.J. (2011). Ultrasound Imaging of Apoptosis: Spectroscopic Detection of DNA-Damage Effects at High and Low Frequencies. In: Didenko, V. (eds) DNA Damage Detection In Situ, Ex Vivo, and In Vivo. Methods in Molecular Biology, vol 682. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-409-8_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-409-8_13

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-408-1

  • Online ISBN: 978-1-60327-409-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation