We have demonstrated a straightforward and noninvasive method to identify the distribution of grana
and starch within an intact leaf. Grana and starch are the major functional structures for
photosynthesis and energy storage of plant, respectively. Both exhibit highly ordered molecular
structures and appear as micrometer-sized granules inside chloroplasts. In order to distinguish grana
and starch, we used multiphoton microscopy, with simultaneous acquisition of two photon fluorescence
(2PF) and second harmonic generation (SHG) signals. Consequently, SHG is found on both grana and
starch while 2PF from chlorophyll indicates the identity of grana.