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Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon
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Focus variation microscope: linear theory and surface tilt sensitivity.

Nikolay Nikolaev, Jon Petzing, Jeremy Coupland

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    |May 4, 2016
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    Summary
    This summary is machine-generated.

    Surface tilt significantly impacts focus variation microscopy (FVM) measurements of microscale roughness. Signal degradation occurs due to mixed specular and diffuse scattering, but can be mitigated with advanced illumination techniques.

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    Area of Science:

    • Surface metrology
    • Optical microscopy
    • Nanotechnology

    Background:

    • Surface roughness measurements are crucial in metrology.
    • Focus variation microscopy (FVM) is a common technique for surface analysis.
    • Previous studies indicated surface tilt influences FVM measurements.

    Purpose of the Study:

    • To investigate the physical principles of image formation in FVM.
    • To analyze the impact of surface tilt on FVM measurements of microscale roughness.
    • To identify methods for mitigating measurement errors caused by surface tilt.

    Main Methods:

    • Theoretical analysis of image formation under the assumption of incoherent scattering.
    • Development of a foil model to simulate surface scattering.
    • Calculation of recorded images using the foil model for various illumination and tilt conditions.

    Main Results:

    • The assumption of incoherent scattering is invalid for microscale roughness with through-the-objective illumination at small tilt angles.
    • Signal degradation and k-space asymmetry occur due to the mixing of specular and diffuse reflection.
    • Extra-aperture illumination or exceeding the aperture acceptance angle can restore the validity of the incoherent assumption.

    Conclusions:

    • Surface tilt introduces systematic errors in FVM measurements of microscale roughness.
    • Ring-light illumination and polarizing optics can prevent or reduce these measurement errors.
    • Careful consideration of illumination and tilt is necessary for accurate FVM surface analysis.