Micro-Brillouin scattering (μ-BR) and a 200 MHz scanning acoustic microscope (SAM) with similar spatial resolutions were applied to evaluate tissue elastic properties in two directions in a trabecula. Acoustic impedance measured by SAM was in the range of 5–9 Mrayl. Wave velocities determined by μ-BR were in the range of (4.75–5.11) × 103 m/s. Both exhibited a similar trend of variation across the trabecula and were significantly correlated (R2 = 0.63–0.67, p < 0.01). μ-BR is useful for the evaluation of tissue stiffness within a trabecula. Combined with SAM or nanoindentation, it can provide additional information to assess elastic anisotropy at the micro-scale.