Bundle sheath (BS) strand cells and BS thylakoids from C4 plants represent a unique system for various studies using a combination of physiological, biochemical, and molecular approaches. We have developed procedures for mechanical disruption of leaf tissues in order to isolate metabolically active bundle sheath strand cells and thylakoids practically free from cross-contamination coming from mesophyll cells. The procedures are described in detail together with useful practical suggestions. Using mechanical disruption we have shown the supramolecular organization of the dimeric LHCII-PSII in BS thylakoids of maize.