<p>The discovery and utilization of novel starches with unique superb properties are highly demanded for modern industrial uses. Small-granule starch (SGS) is a category of unconventional starches with the granular size smaller than 10 μm. The potential use of SGS includes many conventional and novel high-value applications, such as texturizing, fat replacement, encapsulation, controlled delivery and nano-engineering. In the present work, we focused on three SGS isolated from amaranth (<i>Amaranth cruentus</i>), cow cockle (<i>Saponaria vaccaria</i>) and sweet corn (<i>sugary-1</i> maize mutant). The basic structural and unique physical properties of SGS were characterized and compared to common large-granule food starches. It was found that (1) the highly branched amylopectin contributed to low crystallinity and pasting viscosities of sweet corn starch, (2) cow cockle starch exhibited high shear-resistance and low retrogradation in prolonged storage, and (3) the amylopectin for amaranth starch was less branched with small clusters, which was associated with the high crystallinity, medium shear-resistance and low pasting viscosity of amaranth starch. Despite the small size of starch granules, SGS in both native and swelling states showed the capacity of retaining small molecules. Compared to large-granule starch, native SGS are more difficult for small molecules to reach an equilibrium permeation. This work provides insights to the fine structure and physicochemical behaviors of selected high-potent SGS, which is believed to support the industrial production and application of SGS in the future.</p> <p>The characteristics of local polymeric structure dominate many critical properties of glucan particles, such as starch retrogradation and the loading and stabilizing of active substance. Molecular rotor (MR), a fluorescent probe, was proposed to fulfill the simple, high-sensitive, and quantitative-based characterization of local glucan architecture (LGA). In the present work, two innovative studies relevant to this novel method were conducted: (1) MR was able to characterize glucans based on its unique fluorescent response to characteristic LGA, (2) MR was able to sensitively probe and visually demonstrate the transition of LGA induced by starch retrogradation. This novel MR-based approach is expected to advance carbohydrate-related researches in the future.</p>