Dual-channel air-pulse optical coherence elastography for frequency-response analysis

Microliter air-pulse optical coherence elastography (OCE) has recently been proposed for the characterization of soft-tissue biomechanics using transient, sub-nanometer to micrometer-scale natural frequency oscillations. However, previous studies have not been able to provide real-time air-pulse monitoring during OCE natural frequency measurement, which could lead to inaccurate measurement results due to the unknown excitation spectrum. To address this issue, we introduce a dual-channel air-pulse OCE method, with one channel stimulating the sample and the other being simultaneously measured with a pressure sensor. This allows for more accurate natural frequency characterization using the frequency response function, as proven by a comprehensive comparison under different conditions with a diverse range of excitation spectra (from broad to narrow, clean to noisy) as well as a diverse set of sample response spectra. We also demonstrate the capability of the frequency-response analysis in distinguishing samples with different stiffness levels: the dominant natural frequencies increased with agar concentrations (181-359 Hz, concentrations: 1-2%, and maximum displacements: 0.12-0.47 µm) and intraocular pressures (IOPs) for the silicone cornea (333-412 Hz, IOP: 5-40 mmHg, and maximum displacements: 0.41-0.52 µm) under a 200 Pa stimulation pressure. These frequencies remained consistent across different air-pulse durations (3 ms to 35 ms). The dual-channel OCE approach that uses transient, low-pressure stimulation and high-resolution imaging holds the potential to advance our understanding of sample frequency responses, especially when investigating delicate tissues such as the human cornea in vivo.