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Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps.

Publication ,  Journal Article
Battle, C; Lautscham, L; Schmidt, CF
Published in: The Review of scientific instruments
May 2013

Differential interference contrast (DIC) microscopy is a common mode of biological light microscopy used to achieve maximal resolution and contrast with label-free, weakly absorbing specimens such as cells. Maintaining the polarization state of the illuminating light is essential for the technique, and this requirement can conflict with optical trapping. We describe how to optimize DIC imaging using a light-emitting diode illumination source in a microscope while integrating a dual optical trap into the set up. Every time a polarized light beam reflects off or transmits through a dichroic mirror in the beam path, its polarization state will change if it is not polarized exactly parallel (p) or perpendicular (s) to the plane of incidence. We observe wavelength-dependent optical rotation and depolarization effects in our illumination light upon reflection from/transmission through dichroic mirrors in the beam path, resulting in significant degradation of image quality. We describe a method to compensate for these effects by introducing quarter-waveplates and a laser clean-up filter into the imaging pathway. We show that this approach achieves a full recovery of image quality.

Duke Scholars

Published In

The Review of scientific instruments

DOI

EISSN

1089-7623

ISSN

0034-6748

Publication Date

May 2013

Volume

84

Issue

5

Start / End Page

053703

Related Subject Headings

  • Quality Control
  • Optical Tweezers
  • Microscopy, Interference
  • Lighting
  • Applied Physics
  • 51 Physical sciences
  • 40 Engineering
  • 34 Chemical sciences
  • 09 Engineering
  • 03 Chemical Sciences
 

Citation

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Battle, C., Lautscham, L., & Schmidt, C. F. (2013). Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps. The Review of Scientific Instruments, 84(5), 053703. https://doi.org/10.1063/1.4804597
Battle, C., L. Lautscham, and C. F. Schmidt. “Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps.The Review of Scientific Instruments 84, no. 5 (May 2013): 053703. https://doi.org/10.1063/1.4804597.
Battle C, Lautscham L, Schmidt CF. Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps. The Review of scientific instruments. 2013 May;84(5):053703.
Battle, C., et al. “Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps.The Review of Scientific Instruments, vol. 84, no. 5, May 2013, p. 053703. Epmc, doi:10.1063/1.4804597.
Battle C, Lautscham L, Schmidt CF. Differential interference contrast microscopy using light-emitting diode illumination in conjunction with dual optical traps. The Review of scientific instruments. 2013 May;84(5):053703.

Published In

The Review of scientific instruments

DOI

EISSN

1089-7623

ISSN

0034-6748

Publication Date

May 2013

Volume

84

Issue

5

Start / End Page

053703

Related Subject Headings

  • Quality Control
  • Optical Tweezers
  • Microscopy, Interference
  • Lighting
  • Applied Physics
  • 51 Physical sciences
  • 40 Engineering
  • 34 Chemical sciences
  • 09 Engineering
  • 03 Chemical Sciences