14 April 2012 Simultaneous vibration and high-speed microscopy to study mechanotransduction in living cells
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Abstract
Cells exhibit the ability to sense and respond to local mechanical stimuli, leading to changes in function. This capability, referred to as mechanotransduction, is essential to normal tissue function, but the exact mechanisms by which cells sense local forces (strain, shear, compression and vibration) remain unclear. Recent studies in small animals and humans indicate that the frequency of cyclic mechanical stimuli is important, with physiological responses observed for stimuli ranging between 1 and 90 Hz. To better understand the cellular and molecular mechanisms underlying mechanotransduction, it will be important to observe cells in real time, using optical microscopy during high-frequency mechanical stimulation. We have developed a motion-control platform that can produce sinusoidal vibration of live cells during simultaneous high-speed microscopy and fluorimetry, at frequencies up to 100 Hz with peak acceleration up to 9.8 m s-2. The platform is driven by a voice coil and acceleration is measured with an accelerometer (Dytran 7521A1). The motion waveform was verified by high-speed imaging, using a digital camera (Casio EX-F1) operating at 1200 frames s-1 attached to an inverted microscope (Nikon Diaphot). When operating at 45 Hz and 2.94 m s-2 peak acceleration, the observed motion waveform exhibited sinusoidal behaviour, with measured peak-to-peak amplitude of 72 μm. Cultured osteoblast-like cells (UMR-106) were subjected to 2.94 m s-2 vibration at 45 Hz and remained attached and viable. This device provides - for the first time - the capability to mechanically stimulate living cells while simultaneously observing responses with optical microscopy.
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David W. Holdsworth, Hristo N. Nikolov, Jen Au, Kim Beaucage, Jessica Kishimoto, S. Jeffrey Dixon, "Simultaneous vibration and high-speed microscopy to study mechanotransduction in living cells", Proc. SPIE 8317, Medical Imaging 2012: Biomedical Applications in Molecular, Structural, and Functional Imaging, 831715 (14 April 2012); doi: 10.1117/12.913312; https://doi.org/10.1117/12.913312
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KEYWORDS
Microscopy

Optical microscopy

Calcium

Microscopes

Prototyping

Photometry

Tissues

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