Miniaturization and defocus correction for objective-coupled planar illumination microscopy

Opt Lett. 2008 Oct 15;33(20):2302-4. doi: 10.1364/ol.33.002302.


Recently, a light sheet-based technique called objective-coupled planar illumination (OCPI) microscopy [Holekamp et al., Neuron 57, 661 (2008)] was shown to permit low-phototoxicity, high-speed, three-dimensional fluorescence imaging of extended tissue samples. Here, we introduce two major improvements in OCPI microscopy. First, we miniaturize the objective coupler by using a uniaxial gradient-index lens to produce the light sheet. Second, we demonstrate theoretically and experimentally that refractive index mismatch at the fluid/tissue interface causes a significant defocus aberration. By introducing the ability to tune the angle of the light sheet, we show that defocus correction in a miniaturized OCPI microscope leads to a significant improvement in image sharpness deeper into tissue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calibration
  • Diagnostic Imaging / methods
  • Dimethylpolysiloxanes / chemistry
  • Equipment Design
  • Green Fluorescent Proteins / metabolism
  • Light
  • Mice
  • Microscopy / instrumentation*
  • Microscopy / methods
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods
  • Miniaturization*
  • Models, Statistical
  • Olfactory Bulb / pathology
  • Water / chemistry


  • Dimethylpolysiloxanes
  • Water
  • Green Fluorescent Proteins
  • baysilon