Multiphoton excitation-induced photoacoustic microscopy (MEPAM) can be used to investigate the interior of dense
objects precisely and directly because the multiphoton excitation occurs only at the focal point. This method makes it
possible to avoid the strong signal from the surface of dense objects. However, in the case of tissue imaging, one-photon
photoacoustic signals affect the image constructed from MEPAM signals, owing to the smaller cross section of
multiphoton absorption compared with that of one-photon absorption. Thus, in order to apply MEPAM for precise
investigation in living tissues, it is important to enhance (or extract) only the photoacoustic signals induced by
multiphoton excitation.
In this study, we examined the use of frequency-selective detection (frequency filtering) in multiphotonphotoacoustic
imaging by evaluating the depth discrimination and penetration. Because MEPAM signals are generated in
a very small region, they include higher frequency components compared with one-photon photoacoustic signals. We
measured the images at the cross sections of blood-vessel phantoms visualized by MEPAM using the high-frequency
components. We found that the images visualized using only
high-frequency components showed better contrast
compared with those visualized using all frequency components. We conclude that the combination of frequency
filtering and MEPAM demonstrates great potential for precise observation of cross sections of blood vessels in living
tissues.
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