Pupil engineering for extended depth-of-field imaging in a fluorescence miniscope
J. E. Greene, Yujia Xue, Jeffrey Alido, Alex Matlock, Guorong Hu, Kıvılcım Kılıç, Ian G. Davison, Lei Tian
Abstract
SignificanceFluorescence head-mounted microscopes, i.e., miniscopes, have emerged as powerful tools to analyze in-vivo neural populations but exhibit a limited depth-of-field (DoF) due to the use of high numerical aperture (NA) gradient refractive index (GRIN) objective lenses.AimWe present extended depth-of-field (EDoF) miniscope, which integrates an optimized thin and lightweight binary diffractive optical element (DOE) onto the GRIN lens of a miniscope to extend the DoF by 2.8 × between twin foci in fixed scattering samples.ApproachWe use a genetic algorithm that considers the GRIN lens’ aberration and intensity loss from scattering in a Fourier optics-forward model to optimize a DOE and manufacture the DOE through single-step photolithography. We integrate the DOE into EDoF-Miniscope with a lateral accuracy of 70 μm to produce high-contrast signals without compromising the speed, spatial resolution, size, or weight.ResultsWe characterize the performance of EDoF-Miniscope across 5- and 10-μm fluorescent beads embedded in scattering phantoms and demonstrate that EDoF-Miniscope facilitates deeper interrogations of neuronal populations in a 100-μm-thick mouse brain sample and vessels in a whole mouse brain sample.ConclusionsBuilt from off-the-shelf components and augmented by a customizable DOE, we expect that this low-cost EDoF-Miniscope may find utility in a wide range of neural recording applications.