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Photo-imprint Photoacoustic Microscopy for Three-dimensional Label-free Sub-diffraction Imaging
  • Published Date:
    Jan 10 2014
  • Source:
    Phys Rev Lett. 112(1):014302.
Filetype[PDF - 1.77 MB]


Details:
  • Pubmed ID:
    24483902
  • Pubmed Central ID:
    PMC3957272
  • Description:
    Subdiffraction optical microscopy allows the imaging of cellular and subcellular structures with a resolution finer than the diffraction limit. Here, combining the absorption-based photoacoustic effect and intensity-dependent photobleaching effect, we demonstrate a simple method for subdiffraction photoacoustic imaging of both fluorescent and nonfluorescent samples. Our method is based on a double-excitation process, where the first excitation pulse partially and inhomogeneously bleaches the molecules in the diffraction-limited excitation volume, thus biasing the signal contributions from a second excitation pulse striking the same region. The differential signal between the two excitations preserves the signal contribution mostly from the center of the excitation volume, and dramatically sharpens the lateral resolution. Moreover, due to the nonlinear nature of the signal, our method offers an inherent optical sectioning capability, which is lacking in conventional photoacoustic microscopy. By scanning the excitation beam, we performed three-dimensional subdiffraction imaging of varied fluorescent and nonfluorescent species. As any molecules have absorption, this technique has the potential to enable label-free subdiffraction imaging, and can be transferred to other optical imaging modalities or combined with other subdiffraction methods.

  • Document Type:
  • Collection(s):
  • Funding:
    DP1 EB016986/DP/NCCDPHP CDC HHS/United States
    DP1 EB016986/EB/NIBIB NIH HHS/United States
    R01 CA134539/CA/NCI NIH HHS/United States
    R01 CA134539/CA/NCI NIH HHS/United States
    R01 CA157277/CA/NCI NIH HHS/United States
    R01 CA157277/CA/NCI NIH HHS/United States
    R01 CA159959/CA/NCI NIH HHS/United States
    R01 CA159959/CA/NCI NIH HHS/United States
    R01 EB008085/EB/NIBIB NIH HHS/United States
    R01 EB008085/EB/NIBIB NIH HHS/United States
    U54 CA136398/CA/NCI NIH HHS/United States
    U54 CA136398/CA/NCI NIH HHS/United States
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