The present study aimed to extract and isolate cellulose nanocrystals from Ayous (Triplochiton scleroxylon) waste sawdust. Cellulose from Ayous sawdust was obtained by sequential multi-stage chemical extraction and bleaching. After the cellulose extraction, high quality of nanocrystals of cellulose were then extracted by using chemical acid hydrolysis. The chemical compositions, and morphology of nanocrystal of cellulose were analyzed by using standard methods, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), Scanning electron microscopy (SEM) and crystallinity index was determined. A cellulose content of 46.6% was obtained after sequential extraction from the sawdust. SEM images showed the rodlike shape of NCC. According to XRD results, the crystallinity index increased upon chemical extraction. The crystalline index of Ayous wood fiber is 62%, 68% for mercerized cellulose and 86% for nanocrystalline cellulose.