Apparent attenuation by opto-acoustic defocus in phonon microscopy.

Understanding the mechanical properties of biological cells is a challenging problem for the life sciences partly because there are limited methods for mapping elasticity with high resolution. Phonon microscopy is a form of Brillouin light scattering which uses coherent phonons for imaging with elasticity-related contrast, phonon resolution and without labels. It can measure material properties such as sound velocity, acoustic impedance and attenuation. To use it as a contrast mechanism in microscopy, high numerical aperture (NA) lenses are key to high resolution. However, increasing NA induces apparent attenuation, a premature decay of the detected signal. To reduce signal decay and quantify the sound attenuation coefficient in cells, it is necessary to understand the mechanisms that affect signal decay. Here we define opto-acoustic defocus as a signal decay mechanism and propose methods to achieve quantitative sound attenuation measurements, and to optimise in-depth imaging at high resolution which is crucial for cell imaging.

Observing backfolded and unfolded acoustic phonons by broadband optical light scattering.

We use broadband time domain Brillouin scattering to observe coherently generated phonon modes in bulk and nanolayered samples. We transform the measured transients into a frequency-wavevector diagram and compare the resulting dispersion relations to calculations. The detected oscillation amplitude depends on the occupation of phonon modes induced by the pump pulse. For nanolayered samples with an appropriately large period, the whole wavevector range of the Brillouin zone becomes observable by broadband optical light scattering. The backfolded modes vanish, when the excitation has passed the nanolayers and propagates through the substrate underneath.