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Adaptive phase contrast microscopy to compensate for the meniscus effect.

Florian Nienhaus1, Tobias Piotrowski2, Bastian Nießing2

  • 1Fraunhofer Institute for Production Technology IPT, Aachen, Germany. florian.nienhaus@ipt.fraunhofer.de.

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|April 8, 2023
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Summary

Adaptive phase contrast microscopy overcomes liquid lens limitations in cell culture plates. This method significantly expands the observable area for transparent cell observation, especially in small wells.

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Area of Science:

  • Microscopy
  • Cell Biology
  • Optical Engineering

Background:

  • Phase contrast microscopy is crucial for visualizing unstained cells.
  • Cell cultures in microtiter plates create a liquid lens effect due to surface tension.
  • This meniscus effect hinders phase contrast imaging at the curved edges of wells.

Purpose of the Study:

  • To develop an adaptive phase contrast microscopy technique.
  • To optically compensate for the meniscus effect in microtiter plates.
  • To increase the observable area for cell culture analysis.

Main Methods:

  • Replaced the standard condenser annulus with a transmissive liquid crystal display (LCD).
  • Incorporated a deformable, liquid-filled prism in the illumination path.
  • Utilized algorithms to dynamically adjust the LCD and prism based on phase ring displacement.

Main Results:

  • Achieved optical compensation for the liquid lens meniscus effect.
  • Enabled simultaneous observation of phase contrast images and optical path adjustments.
  • Demonstrated a significant increase in observable area, over twelvefold for 96-well plates.

Conclusions:

  • Adaptive phase contrast microscopy enhances cell observation in microtiter plates.
  • The technique effectively overcomes limitations posed by liquid lenses in well-based cultures.
  • This advancement offers greater efficiency for examining cell cultures in microtiter plate formats.