A non-destructive methodology for monitoring impedance changes in sorghum leaves was developed and recorded irrigation-dependent responses that differed between leaf tissues. Metal microneedles were used as impedance probes and were shown to cause minimal damage to the plant. The needles were placed on either the abaxial or adaxial side of the leaf midrib using small clamps and re-used hundreds of times with minimal signs of wear. Cross-sectional images verified the precision of microneedle placement near vascular bundles on the abaxial surface and in non-vascular hydrenchyma on the adaxial surface. Impedance measurements with microneedles displayed a significant decrease in resistance compared to planar electrodes due to bypassing the epidermal layer. A tissue-specific impedance response was seen in relation to irrigation where the non-vascular adaxial surface remained largely stable throughout a day of measurement, while impedance increased in the vascular abaxial surface during exposure to light and decreased following watering. Impedance data were also compared with simultaneous gas exchange measurements of photosynthesis and transpiration.