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Micro- and nano-platelets are a group of particles with typical flat surfaces and hexagonal shape. They are obtained by hierarchical self-assembly in water of α-cyclodextrin and polysaccharides hydrophobically-modified with alkyl chains. It is expected that the formation of well structured and cohesive platelets is driven by the interaction between alkyl chains grafted on polysaccharides and α-cyclodextrin. The objective of this investigation is to tune platelet formation by modifying these two parameters, independently on polysaccharide composition. A systematic study was conducted by varying polysaccharide type (dextran, pullulan, amylopectin), degree of substitution (DS:0.1–5.6%) and α-cyclodextrin concentration (0–10wt%) for a fixed concentration of polysaccharide esterified with palmitoyl groups (1wt%). Characterizations include ATR-FTIR, elemental analysis, solid state 13C NMR and transmission electron microscopy. Abundant and well-organized hexagonal platelets were obtained with high DS (4.2–5.6%) and a concentration of α-cyclodextrin higher than 2.5wt%. Isothermal titration calorimetry revealed a sequential binding with a stoichiometry of 2 α-cyclodextrin molecules for 1 palmitoyl group grafted on dextran. This is the first report showing the possibility to control platelet formation by modifying DS and α-cyclodextrin concentration, independently on polysaccharide composition.