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Proceedings Paper

The use of two-photon microscopy to study the biological effects of focused ultrasound on the brain
Author(s): Alison Burgess; Eunice E. Cho; Leila Shaffaf; Tam Nhan; Charissa Poon; Kullervo Hynynen
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Paper Abstract

Focused ultrasound (FUS) has been used to successfully disrupt the blood-brain barrier (BBB), aiding in the delivery of therapeutic agents to the brain and leading to improvements in disease pathology. Although significant progress has been made in the development of FUS technology, there is still a lack of understanding of the biophysical mechanisms of the BBB disruption and the microscopic effects of this disruption on brain cells. In this study, we combine a custom built ultrasound transducer with two-photon microscopy to conduct real time monitoring of BBB disruption in vivo. We have manufactured and tested a single element piezoelectric transducer with frequencies ranging from 1.15 to 1.30 MHz. Sonications were performed using 0.07-0.25 MPa estimated in situ pressure, 10 ms pulses, 1 Hz pulse repetition frequency for a total duration of 120 s in the presence of microbubbles. BBB disruption was observed through a cranial window created in the rat skull after intravenous injection of dextran conjugated- Texas Red (MW: 10,000 - 70,000 Da). Using this experimental setup, we have observed and characterized 3 different leakage patterns following BBB disruption. Our results indicate that varying the acoustic power leading to in situ pressure changes, may allow us to control the mechanism of BBB disruption. Furthermore, we have labelled astrocytes in vivo in order to visualize the effects of FUS on this cell population. Combination of our custom transducers with two-photon microscopy will allow significant advancement in allow significant advancement in the understanding of the mechanisms and cellular effects of FUS-induced BBB disruption.

Paper Details

Date Published: 9 February 2012
PDF: 7 pages
Proc. SPIE 8226, Multiphoton Microscopy in the Biomedical Sciences XII, 822642 (9 February 2012); doi: 10.1117/12.908826
Show Author Affiliations
Alison Burgess, Sunnybrook Health Sciences Ctr. (Canada)
Eunice E. Cho, Sunnybrook Health Sciences Ctr. (Canada)
Leila Shaffaf, Sunnybrook Health Sciences Ctr. (Canada)
Univ. of Toronto (Canada)
Tam Nhan, Sunnybrook Health Sciences Ctr. (Canada)
Univ. of Toronto (Canada)
Charissa Poon, Sunnybrook Health Sciences Ctr. (Canada)
Kullervo Hynynen, Sunnybrook Health Sciences Ctr. (Canada)
Univ. of Toronto (Canada)

Published in SPIE Proceedings Vol. 8226:
Multiphoton Microscopy in the Biomedical Sciences XII
Karsten König, Editor(s)

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