3D imaging of biological specimens using the LUCID clearing reagent and the HandySPIM lens-free light-sheet module

We previously reported the tissue-clearing reagent LUCID and the lens-free LED light-sheet microscope module HandySPIM. In the present study, we further advance both technologies and demonstrate that LUCID exhibits high compatibility with immunostaining, nuclear staining, and fluorescent protein labeling. In parallel, HandySPIM was upgraded to support selectable LED excitation wavelengths across the visible spectrum (400–600 nm) and automated image acquisition along the vertical (Z) axis. As a result, the integrated use of LUCID and HandySPIM enables simple, low-cost, three-dimensional fluorescence imaging using conventional two-dimensional microscopes.