Journal: bioRxiv

Article Title: Polarization-engineered aberration-resilient light sheet microscopy

doi: 10.64898/2026.05.11.724351

Figure Lengend Snippet: (A) Optical layout of the beam shaping unit of PEARLS. HWP, half-wave plate; VBE, variable beam expander; PBS, polarizing beamsplitter; BE, beam expander; PL, Powell lens; IR, iris; CL, cylindrical lens; L, lens; QWP, quarter-wave plate; FBP, Fresnel biprism; M, mirror. (B) Demonstrations of PEARLS’ tunability of axial resolution (i-iii, NA max /NA min =0.46/0.38, 0.6/0.52, and 0.36/0.28); confinement (iv-v, NA max /NA min =0.46/0.42 and 0.5/0.38); optical transfer function (OTF) weighting (NA max /NA min =0.46/0.38, intensity ratio of outer to inner CSW on the rear pupil plane is 1:1, 4:1, 1:2 for green (i), blue (vi), and orange (vii), respectively). Arrows in (viii) indicate the axial spatial frequency contributions from the individual CSW components (black: outer CSW; gray: inner CSW); and light sheet types (i, hexagonal-type, ix, square-type, x, CSW, and xi, a profile without an equivalent LLS). Simulations (left) and experiments (right) show good agreement. Scale bar: 5-λ exc /n. (C) PEARLS light sheet profiles (hexagonal type, NA max /NA min =0.6/0.5) at six different wavelengths, ranging from 445, 488, 514, 560, 607, and 637nm. Scale bar: 2-μm. (D) Relationship between the wavelength and the corresponding axial period. Linear fitting yields R ² = 0.99. (E) Photobleaching comparisons of hexagonal type PEARLS and dithered LLS with identical profile (Video S3). Each frame was normalized with respect to (w.r.t.) the maximum intensity of the Frame 1 of each light sheet. Representative Frames 1, 600, 1500, and 4000 taken by LLS (top) and PEARLS (bottom) were shown. Scale bar: 5-μm. (F) Averaged photobleaching curves of PEARLS (blue) and LLS (red). The shaded areas indicate ±1 standard deviation (SD) from five datasets taken at five different regions of the same collagen gel with intensities normalized. Inset: characteristic decay constants of LLS (red) and PEARLS (blue) obtained by fitting individual photobleaching curves with exponential decays. Horizontal and vertical lines indicate the mean and ±1 SD of the five datasets, respectively. (G) Comparison of PEARLS and LLS applied to imaging within a microfluidic device. (H) Transverse cross-section profiles and overall XZ PSFs of LLS (left) and PEARLS (right) without (top) and with (bottom) the microfluidic device. Scale bars: 1-μm. (I) XZ views of raw collagen images within the microfluidic device taken by LLS (top) and PEARLS (bottom) with intensity normalized w.r.t. the maximum intensities of each volume. Scale bar: 5-μm. Red and blue curves show intensities of the corresponding line cuts. Raw collagen images are shown and used for analysis in (E, F, I).

Article Snippet: Lastly, the light sheet beam enters the third module to generate the CSWs: a third polarizing beamsplitter cube (Thorlabs, PBS251) splits the beam again into orthogonal polarization paths—each path is composed of an achromatic quarter-wave plate (Bolder Vision Optik AHWP3)—to form an optical insulator, a Fresnel biprism (170-degree, Newlight Photonics FBP2020G-170) with custom antireflective coating that splits the light in z-direction, and a mirror.

Techniques: Standard Deviation, Comparison, Imaging