Diagonally Scanned Light-Sheet Microscopy for Fast Volumetric Imaging of Adherent Cells.

ABSTRACT: In subcellular light-sheet fluorescence microscopy (LSFM) of adherent cells, glass substrates are advantageously rotated relative to the excitation and emission light paths to avoid glass-induced optical aberrations. Because cells are spread across the sample volume, three-dimensional imaging requires a light-sheet with a long propagation length, or rapid sample scanning. However, the former degrades axial resolution and/or optical sectioning, while the latter mechanically perturbs sensitive biological specimens on pliant biomimetic substrates (e.g., collagen and basement membrane). Here, we use aberration-free remote focusing to diagonally sweep a narrow light-sheet along the sample surface, enabling multicolor imaging with high spatiotemporal resolution. Further, we implement a dithered

SUBMITTER: Dean KM 

PROVIDER: S-EPMC4816690 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature