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Updated: Apr 20, 2026

Three-dimensional Imaging of Bacterial Cells for Accurate Cellular Representations and Precise Protein Localization
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Interferometric three-dimensional single molecule localization microscopy using a single high-numerical-aperture

P Zhang, P M Goodwin, J H Werner

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    |November 18, 2014
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    Summary
    This summary is machine-generated.

    We developed a new single-objective interferometric method for super-resolution microscopy. This technique achieves nanometer axial localization precision, simplifying 3D super-resolution imaging and reducing system costs.

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    Area of Science:

    • Biophysics
    • Optical Microscopy
    • Nanotechnology

    Background:

    • Interferometric photoactivated localization microscopy (iPALM) offers nanometer axial localization accuracy for 3D super-resolution imaging.
    • Traditional iPALM requires two high-numerical-aperture (NA) objectives, increasing cost and limiting sample compatibility.

    Purpose of the Study:

    • To develop a simplified interferometric single-molecule localization microscopy method using a single high-NA objective.
    • To achieve nanometer precision in axial localization for 3D super-resolution imaging with a more accessible setup.

    Main Methods:

    • Implementation of a novel interferometric single-molecule localization microscopy technique.
    • Utilizing wavefront-splitting interference with a single high-NA objective for fluorescence collection.
    • Analysis of phase-shifted interference signals to determine axial positions.

    Main Results:

    • Demonstrated unambiguous determination of single-molecule axial positions with nanometer precision over a 2λ range.
    • Simplified system configuration and sample mounting compared to dual-objective iPALM.
    • Minimized optical path drift by employing wavefront-splitting interference.

    Conclusions:

    • The proposed single-objective method offers a cost-effective and simplified approach to 3D super-resolution imaging.
    • Achieves high axial localization accuracy, comparable to existing methods.
    • Potential for improved long-term 3D super-resolution imaging due to reduced drift.