Spatio-temporal performance enhancement in an incoherent holography lattice light-sheet microscope (IHLLS)

We propose an incoherent holography detection technique for lattice light-sheet (IHLLS) systems for 3D imaging without moving either the sample stage or the detection microscope objective, providing intrinsic instrumental simplicity and high accuracy when compared to the original LLS schemes. The approach is based on a modified dual-lens Fresnel Incoherent Correlation Holography technique to produce a complex hologram and to provide the distance needed for the hologram reconstruction. We report an IHLLS microscope, including characterization of the sensor performance, and demonstrate a significant contrast improvement on beads and neuronal structures within a biological test sample as well as quantitative phase imaging. The IHLLS has similar or better transverse performances when compared to the LLS technique. In addition, the IHLLS allows for volume reconstruction from fewer z-galvo displacements, thus facilitating faster volume acquisition. However, the axial performance is slightly degraded for the IHLLS, when moving the z-galvo in 7 or 9 steps along the 60 μm or 80 μm ranges. The axial performances may be improved by increasing the number of the z-galvo steps.