OSTEOBLAST ADHESION OF BREAST CANCER CELLS WITH SCANNING ACOUSTIC MICROSCOPY Page: 2 of 8
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OSTEOBLAST ADHESION OF BREAST CANCER CELLS WITH SCANNING
Department of Physics, Idaho National Engineering and Environmental Laboratory
Idaho Falls, ID 83415, USA
R. R. MERCER and A. M. MASTRO*2
Department of Biochemistry and Molecular Biology, The Pennsylvania State University
431S Frear, University Park, PA 16802, USA
Once breast cancer has progressed to an advanced stage, it is likely to metastasize to bone, and is
usually fatal. However, the process by which breast cancer affects bone tissue is poorly understood 
When breast cancer metastasizes to bone, the osteoclasts are constitutively activated, resulting in osteolytic
lesions . Kureja et al examined nude mice with bone metastasis, and found a significant decrease in
bone formation . Similar results have also been reported elsewhere -. Therefore, these
observations suggest that breast cancer cells affect osteoblasts in addition to osteoclasts.
When MC3T3-E1 osteoblasts were cultured in cancer cell condition medium, they took on a fibroblast-like
morphology and exhibited adhesion characteristics different from those observed in normal medium. The
osteoblasts were fixed and stained for actin visualization and for observation of the focal adhesion plaques
with optical microscopy. However, since fixation kills the cells, some characteristics of the adhesion may
be missed. On the other hand, an acoustic image is formed by reflected ultrasonic waves that are based on
the elastic properties of the living cells. Therefore, fixing and staining are not required for mechanical
scanning acoustic reflection microscopy  (hereinafter called simply "SAM"). Hence, living cells can be
easily observed. Further, SAM allows observation not only of the surface but also of the internal structure
of the specimen with sub-micrometer resolution -. This report presents the visualization of adhesive
conditions of living osteoblasts grown on the substrate using SAM. The results are compared with those
obtained with laser scanning confocal microscopy.
2. Principle of Acoustic Imaging
Figure 1 is the schematic diagram of the SAM. Referring to Fig. 1, the imaging principle of SAM is
described as follows:
The SAM instrument comprises a transmitting/receiving section, an X-Y scanning section, a Z
scanning section, a computer section for controlling the SAM, and a display section for observing a
specimen. The transmitting/receiving section includes a transmitter, a receiver having an amplifier, and a
circulator. The X-Y scanning section comprises an X-Y stage including a temperature-controlled chamber
for containing living cells grown on a substrate in a coupling medium (i.e., a culture liquid). The Z
scanning section includes a Z-stage and an acoustic lens. An electrical signal (i.e., tone-burst wave)
generated by a transmitter inputs to a piezoelectric transducer (i.e., zinc oxide), located on the top of a
buffer rod through a circulator. The input voltage from the transmitter to the transducer is approximately
5V. The electrical signal is converted into an acoustic signal (i.e., ultrasonic plane wave) by the transducer.
The ultrasonic plane wave travels through the buffer rod made of sapphire to a spherical recess (hereinafter
*1 E-mail: mivascrinel.gov, Telephone: 208-526-8951, Facsimile: 208-526-0690
*2 E-mail: a36Apsu.edu, Telephone: 814-863-0152, Facsimile: 814-863-7024
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Miyasaka, Chiaki; Mercer, Robyn R.; Mastro, Andrea M. & Telschow, Ken L. OSTEOBLAST ADHESION OF BREAST CANCER CELLS WITH SCANNING ACOUSTIC MICROSCOPY, article, March 1, 2005; [Idaho Falls, Idaho]. (digital.library.unt.edu/ark:/67531/metadc879107/m1/2/: accessed February 21, 2019), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.