Miniaturization and defocus correction for objective-coupled planar illumination microscopy

摘要

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