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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
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Accurate position tracking of optically trapped live cells.

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Directly trapping cells with optical tweezers overcomes limitations of bead-based methods. This study models cell roll instabilities and presents novel techniques for stable cell manipulation in life science research.

Keywords:
(070.6120) Spatial light modulators(110.2960) Image analysis(170.0180) Microscopy(180.2520) Fluorescence microscopy(350.4855) Optical tweezers or optical manipulation

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Area of Science:

  • Biophysics
  • Optical Trapping
  • Cellular Mechanics

Background:

  • Optical trapping is crucial in life science, but bead-based methods can alter biological functions.
  • Direct cell trapping is desired, yet instabilities like cell roll hinder precise measurements.
  • These instabilities arise from viscous drag, thermal convection, and Brownian forces.

Purpose of the Study:

  • To mathematically model the causes of cell roll instability during optical trapping.
  • To present experimental approaches for stable manipulation of large, inhomogeneous biological samples.
  • To improve the reliability of measurements in optical trapping experiments.

Main Methods:

  • Mathematical modeling of cell roll and re-orientation dynamics.
  • Development of a novel three-trap beam profile for enhanced stability.
  • Utilizing fluorescence imaging analysis for precise tracking of trapped cells.

Main Results:

  • Successfully modeled the physical forces causing cell roll in optical traps.
  • Demonstrated improved trapping stability using a multi-trap configuration.
  • Showcased the efficacy of fluorescence imaging for accurate cell tracking.

Conclusions:

  • Direct cell trapping is feasible and advantageous over bead-based methods.
  • The presented methods effectively mitigate instabilities in trapping large, inhomogeneous cells.
  • These techniques are applicable to various cell types, including T cells, and other challenging samples.