Reflection‐mode switchable subwavelength Bessel‐beam and Gaussian‐beam photoacoustic microscopy in vivo

We have developed a reflection-mode switchable subwavelength Bessel-beam (BB) and Gaussian-beam (GB) photoacoustic microscopy (PAM) system. To achieve both reflection-mode and high resolution, we tightly attached a very small ultrasound transducer to an optical objective lens with numerical aperture of 1.0 and working distance of 2.5 mm. We used axicon and an achromatic doublet in our system to obtain the extended depth of field (DOF) of the BB. To compare the DOF performance achieved with our BB-PAM system against GB-PAM system, we designed our system so that the GB can be easily generated by simply removing the lenses. Using a 532 nm pulse laser, we achieved the lateral resolutions of 300 and 270 nm for BB-PAM and GB-PAM, respectively. The measured DOF of BB-PAM was approximately 229 μm, which was about 7× better than that of GB-PAM. We imaged the vasculature of a mouse ear using BB-PAM and GB-PAM and confirmed that the DOF of BB-PAM is much better than the DOF of GB-PAM. Thus, we believe that the high resolution achieved at the extended DOF by our system is very practical for wide range of biomedical research including red blood cell (RBC) migration in blood vessels at various depths and observation of cell migration or cell culture.

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