In vivo functional photoacoustic micro-imaging of the electrically stimulated rat brain with multiwavelengths

In this study, we report on using multi-wavelength photoacoustic microscopy to image hemodynamic changes of total hemoglobin concentration (HbT) (i.e., blood volume) and oxygenation (SO2) in rat brain cortex vessels with electrical stimulation. Electrical stimulation of the rat left forelimb was applied to evoke changes in vascular dynamics of the rat somatosensory cortex. The applied current pulses were with a pulse frequency of 3 Hz, pulse duration of 0.2 ms, and pulse amplitude of 5 mA, respectively. The imaging target of rat brains was demarcated at AP 0 - -2.5 mm and ML ± 6 mm with respect to bregma. HbT changes were probed by images acquired at 570 nm, a hemoglobin isosbestic point while SO2 changes were imaged by those acquired at 560 nm or 600 nm and their derivatives, which were normalized to those with 570 nm wavelengths. Correlation between the electrical stimulation paradigm and images acquired at 570, 560, and 600 nm in contralateral and ipsilateral vasculature was statistically analyzed, showing that the HbT and SO2 changes revealed by multi-wavelength photoacoustic images spatially correlated with contralateral vasculature.