| First Author | Silvestri L | Year | 2021 |
| Journal | Nat Methods | Volume | 18 |
| Issue | 8 | Pages | 953-958 |
| PubMed ID | 34312564 | Mgi Jnum | J:359583 |
| Mgi Id | MGI:7788548 | Doi | 10.1038/s41592-021-01208-1 |
| Citation | Silvestri L, et al. (2021) Universal autofocus for quantitative volumetric microscopy of whole mouse brains. Nat Methods 18(8):953-958 |
| abstractText | Unbiased quantitative analysis of macroscopic biological samples demands fast imaging systems capable of maintaining high resolution across large volumes. Here we introduce RAPID (rapid autofocusing via pupil-split image phase detection), a real-time autofocus method applicable in every widefield-based microscope. RAPID-enabled light-sheet microscopy reliably reconstructs intact, cleared mouse brains with subcellular resolution, and allowed us to characterize the three-dimensional (3D) spatial clustering of somatostatin-positive neurons in the whole encephalon, including densely labeled areas. Furthermore, it enabled 3D morphological analysis of microglia across the entire brain. Beyond light-sheet microscopy, we demonstrate that RAPID maintains high image quality in various settings, from in vivo fluorescence imaging to 3D tracking of fast-moving organisms. RAPID thus provides a flexible autofocus solution that is suitable for traditional automated microscopy tasks as well as for quantitative analysis of large biological specimens. |
