This study aimed to explore the effect of various noise definition criteria in linear extrapolation technique to noise level baseline on steady-state visual evoked potential (SSVEP)-based visual acuity assessment. Four noise definition criteria on frequency-domain, i.e., the mean amplitude at the two adjacent bins of the target frequency, the mean amplitude of a narrow frequency band on either side of the target frequency, the mean amplitude at a broad frequency band except for the target frequency and its harmonic frequencies, and the mean amplitude at a broad frequency band at resting state, corresponding to noise 1, noise 2, noise 3, and noise 4, were introduced to calculate noise level baselines. Then, two experiments were implemented. In experiment 1, electroencephalography (EEG) signals of resting state were recorded for fourteen subjects. In experiment 2, the visual stimuli of vertical sinusoidal gratings at six spatial frequency steps were used to induce SSVEPs for twelve subjects. Finally, SSVEP visual acuity was obtained via the SSVEP visual acuity threshold estimation of linear extrapolation technique to noise level baseline with various noise definition criteria. The bland-Altman analysis found that the difference between subjective Freiburg Visual Acuity and Contrast Test (FrACT) and objective SSVEP visual acuity was - 0.0892, - 0.1071, - 0.0745, and - 0.0804 logMAR and the 95% limit of agreement was 0.2150, 0.2146, 0.2046, and 0.2189 logMAR for noise 1, noise 2, noise 3, and noise 4, respectively, indicating that visual acuity of noise 3 definition criterion, i.e., the mean amplitude at a broad frequency band except for the target frequency and its harmonic frequencies, showed the best performance. This study recommended noise definition criterion 3 of the mean amplitude at a broad frequency band to calculate the noise level baseline in the linear extrapolation of SSVEP-based visual acuity assessment.