Frequency-Dependent Bioimpedance Signatures of Ocular Tissues in Intact Ex Vivo Eyes Under Simulated Surgical Conditions.

Purpose: To characterize the frequency-dependent bioimpedance properties of major ocular tissues in intact ex vivo porcine eyes under simulated surgical conditions and evaluate tissue separability at discrete frequencies. Methods: Bioimpedance spectra were acquired from sclera, corneal epithelium, iris, lens, vitreous, and retina in intact ex vivo porcine eyes using a two-electrode probe and a precision LCR meter over 5 kHz to 1 MHz. Measurements were obtained under balanced salt solution and ophthalmic viscosurgical device conditions. Probe-tissue contact was verified by microscope visualization and optical coherence tomography. Tissue separability at 5, 50, 100, and 900 kHz was evaluated using global and pairwise statistical comparisons, effect sizes, and ROC-based separability metrics. Robotic-stabilized and handheld measurements were also compared. Results: Ocular tissues demonstrated distinct, frequency-dependent impedance magnitude distributions. Across sampled frequencies, 60% to 80% of tissue pairs showed significant differences after multiplicity correction. Median pairwise effect sizes ranged from Cohen's d = 0.48 at 5 kHz to 1.04 to 1.06 at 50 to 100 kHz. Median ROC-based separability was 0.91 at 5 kHz and 0.76 to 0.77 at 50 to 900 kHz. Robotic-stabilized measurements showed lower variance than handheld measurements, although tissue-specific impedance ranges and frequency-dependent trends were preserved across acquisition modes. Conclusions: Major ocular tissues exhibit reproducible, frequency-dependent bioimpedance signatures in intact ex vivo eyes under simulated surgical preparation. These findings establish a physiologically relevant ocular impedance reference dataset and support bioimpedance as a complementary modality for tissue differentiation in ophthalmic microsurgery.