A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues.
Adam K GlaserKevin W BishopLindsey A BarnerEtsuo A SusakiShimpei I KubotaGan GaoRobert B SerafinPooja BalaramEmily TurschakPhilip R NicovichHoyin LaiLuciano A G LucasYating YiEva K NicholsHongyi HuangNicholas P RederJasmine J WilsonRamya SivakumarElya ShamskhouCaleb R StoltzfusXing WeiAndrew K HemptonMarko PendePrayag MurawalaHans-Ulrich DodtTakato ImaizumiJay ShendureBrian J BeliveauMichael Y GernerLi XinHu ZhaoLawrence D TrueR Clay ReidJayaram ChandrashekarHiroki R UedaKarel SvobodaJonathan T C LiuPublished in: Nature methods (2022)
Light-sheet microscopy has emerged as the preferred means for high-throughput volumetric imaging of cleared tissues. However, there is a need for a flexible system that can address imaging applications with varied requirements in terms of resolution, sample size, tissue-clearing protocol, and transparent sample-holder material. Here, we present a 'hybrid' system that combines a unique non-orthogonal dual-objective and conventional (orthogonal) open-top light-sheet (OTLS) architecture for versatile multi-scale volumetric imaging. We demonstrate efficient screening and targeted sub-micrometer imaging of sparse axons within an intact, cleared mouse brain. The same system enables high-throughput automated imaging of multiple specimens, as spotlighted by a quantitative multi-scale analysis of brain metastases. Compared with existing academic and commercial light-sheet microscopy systems, our hybrid OTLS system provides a unique combination of versatility and performance necessary to satisfy the diverse requirements of a growing number of cleared-tissue imaging applications.