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This study introduces optical focal plane shifting for depth scanning in photoacoustic remote sensing microscopy (PARS). This non-contact method enables label-free imaging of biological structures without mechanical scanning.

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Biophotonics

Background:

  • Conventional microscopy often requires mechanical scanning for depth imaging, which can be detrimental to delicate biological samples.
  • Photoacoustic Remote Sensing Microscopy (PARS) offers label-free imaging capabilities but traditionally lacks efficient axial scanning.
  • Optical focal plane shifting presents a non-invasive alternative for depth visualization in biological imaging.

Purpose of the Study:

  • To develop and demonstrate an optical method for axial scanning in PARS.
  • To enable depth imaging of biological structures without physical sample manipulation.
  • To advance label-free, non-contact microscopy techniques for biological research.

Main Methods:

  • Integration of a deformable mirror as a varifocal element within a PARS system.
  • Implementation of optical focal plane shifting for axial scanning (Δz ∼ 240 µm).
  • System validation using USAF resolution targets and carbon fiber phantoms, followed by in-vivo imaging.

Main Results:

  • Demonstrated successful optical focal plane shifting for depth scanning.
  • Achieved axial scanning range of approximately 240 µm.
  • Successfully visualized in-vivo blood vessels in chicken embryo chorioallantoic membrane (CAM).

Conclusions:

  • The deformable mirror-based PARS system enables non-contact axial scanning.
  • This technique is a promising step towards aberration-free, label-free PARS imaging with depth visualization.
  • Further development could lead to advanced non-invasive imaging tools for delicate biological samples.