Purpose: To evaluate and compare the temporal changes in pulse waveform parameters of ocular blood flow (OBF) between non-habitual and habitual groups due to caffeine intake. Method: This study was conducted on 19 healthy subjects (non-habitual 8; habitual 11), non-smoking and between 21 and 30 years of age. Using laser speckle flowgraphy (LSFG), three areas of optical nerve head were analyzed which are vessel, tissue, and overall, each with ten pulse waveform parameters, namely mean blur rate (MBR), fluctuation, skew, blowout score (BOS), blowout time (BOT), rising rate, falling rate, flow acceleration index (FAI), acceleration time index (ATI), and resistive index (RI). Two-way mixed ANOVA was used to determine the difference between every two groups where p < 0.05 is considered significant. Result: There were significant differences between the two groups in several ocular pulse waveform parameters, namely MBR (overall, vessel, tissue), BOT (overall), rising rate (overall), and falling rate (vessel), all with p < 0.05. In addition, the ocular pulse waveform parameters, i.e., MBR (overall), skew (tissue), and BOT (tissue) showed significant temporal changes within the non-habitual group, but not within the habitual group. The temporal changes in parameters MBR (vessel, tissue), skew (overall, vessel), BOT (overall, vessel), rising rate (overall), falling rate (overall, vessel), and FAI (tissue) were significant for both groups (habitual and non-habitual) in response to caffeine intake. Conclusion: The experiment results demonstrated caffeine does modulate OBF significantly and response differently in non-habitual and habitual groups. Among all ten parameters, MBR and BOT were identified as the suitable biomarkers to differentiate between the two groups. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.