Despite the enormous potential of cellulose nanofibrils (CNFs) as a reinforcing filler in various fields, the use of them has been limited by high-energy mechanical treatments that require a lot of energy and time consumption. To reduce the demands of energy and time required for mechanical treatments, microalgae, in particular, Nannochloropsis oceanica, which has small size, rapid growth rate, and high productivity was used as a CNFs source. This study obtains the CNFs by lipid/protein extraction, purification, and TEMPO-mediated oxidation processes under gentle mixing without high-energy mechanical treatments. Furthermore, to evaluate the applicability of microalgal CNFs as a reinforcing filler, this study estimated the mechanical strength of the fibrils by the sonication-induced scission method. To achieve a precise estimation, an effective method to distinguish straight fibrils from buckled fibrils was also developed, and subsequently, only straight fibrils were used to calculate the mechanical strength in the sonication-induced scission method. Consequently, the tensile strength of the N. oceanica CNFs is around 3–4 GPa on average which is comparable with the mechanical strength of general reinforcing fillers and even higher than that of wood CNFs. Thus, this study has shown that the newly proposed simplified method using N. oceanica is very successful in producing CNFs with great mechanical strength which could be used in various reinforcement fields.