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Significance The initial dip in hemoglobin-oxygenation response to stimulations is a spatially confined endogenous indicator that is faster than the blood flow response, making it a desired label-free contrast to map the neural activity. A fundamental question is whether a single-impulse stimulus, much shorter than the response delay, could produce an observable initial dip without repeated stimulation. Aim To answer this question, we report high-speed functional photoacoustic (PA) microscopy to investigate the initial dip in mouse brain