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Multiview microscopy of single cells through microstructure-based indirect optical manipulation.

Gaszton Vizsnyiczai1,2, András Búzás1,2, Badri Lakshmanrao Aekbote1,3

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Summary

Holographic optical tweezers enable precise rotation of single cells for improved 3D fluorescence imaging. This technique overcomes limited axial resolution in microscopy, achieving isotropic optical resolution for clearer cell visualization.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Cell Biology

Background:

  • Microscopy's axial resolution is limited by optics, causing anisotropic 3D imaging.
  • Existing optical solutions for anisotropic resolution are complex and require high stability.
  • Multiview imaging via sample rotation is a potential solution.

Purpose of the Study:

  • To develop a straightforward method for achieving isotropic optical resolution in 3D cell imaging.
  • To implement multiview imaging of single cells using holographic optical tweezers.

Main Methods:

  • Cells were indirectly trapped and manipulated using microtools fabricated with two-photon polymerization.
  • Holographic optical tweezers rotated cells around an axis perpendicular to the optical axis.
  • Multiview image processing combined stacks into a single 3D array.

Main Results:

  • Precise 6-degree-of-freedom manipulation of single cells was achieved.
  • Multiview fluorescence imaging from arbitrary directions was enabled.
  • Isotropic optical resolution approaching the lateral diffraction limit was obtained.

Conclusions:

  • Holographic optical tweezers offer a robust method for precise cell manipulation and multiview imaging.
  • This approach effectively eliminates anisotropic resolution in 3D fluorescence microscopy.
  • The technique is adaptable to various microscopy platforms.