title: Low-cost multimodal light sheet microscopy for optically cleared tissues and living specimens.
abstract: Light sheet microscopy techniques have expanded with designs to address many new applications. Due to rapid advancements in computing power, camera/detector technologies, and tissue clearing techniques, light sheet methods are becoming increasingly popular for biomedical imaging applications at the cellular and tissue levels. Light sheet imaging modalities couple rapid imaging rates, low-levels of phototoxicity, and excellent signal to noise ratios, contributing to their popularity for experimental biology. However, the current major limitation of light sheet microscopy arises from optical aberrations, with the main drawback being the defocusing introduced by refractive index variations that accompany clearing techniques. Here, we propose an inexpensive and easy to build light sheet based instrumentation to overcome this limitation by optomechanically decoupling the sample scanning movement from the detection step. Our solution is relatively simple to implement and also provides increased modularity by using a swappable excitation arm. This expands the range of samples we can image on a single system, from high resolution for single cells at ? m spatial resolution, to tissues with mm spatial resolution. We demonstrate our approach, using the system to image iDISCO cleared embryos and sciatic nerves, and provide the full three-dimensional reconstruction of these objects in minutes.
authors.affiliation: McGill University, Department of Chemistry, 801 Sherbrooke Street, West, Montreal, Quebec H3A 0B8, Canada.
authors.forename: Vincent
authors.initials: V
authors.lastname: Rouger
authors.affiliation: McGill University, Montreal Neurological Institute, Department of Neurology, 3801 University Street, Montreal, Quebec H3A 2B4, Canada.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Alchini
authors.affiliation: McGill University, Department of Chemistry, 801 Sherbrooke Street, West, Montreal, Quebec H3A 0B8, Canada.
authors.forename: Alexei
authors.initials: A
authors.lastname: Kazarine
authors.affiliation: McGill University, Department of Chemistry, 801 Sherbrooke Street, West, Montreal, Quebec H3A 0B8, Canada.
authors.forename: Angelica A
authors.initials: AA
authors.lastname: Gopal
authors.affiliation: McGill University, Montreal Neurological Institute, Department of Neurology, 3801 University Street, Montreal, Quebec H3A 2B4, Canada.
authors.forename: Marie-Pier
authors.initials: MP
authors.lastname: Girouard
authors.affiliation: McGill University, Montreal Neurological Institute, Department of Neurology, 3801 University Street, Montreal, Quebec H3A 2B4, Canada.
authors.forename: Alyson E
authors.initials: AE
authors.lastname: Fournier
authors.affiliation: McGill University, Department of Chemistry, 801 Sherbrooke Street, West, Montreal, Quebec H3A 0B8, CanadacMcGill University, Department of Physics, 3600 University Street, Montreal, Quebec H3A 2T8, Canada.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Wiseman
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian / Q000000981:diagnostic imaging; D004867:Equipment Design; D005260:Female; D005727:Ganglia, Spinal / Q000000981:diagnostic imaging; D057809:HEK293 Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007834:Lasers; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D064847:Multimodal Imaging / Q000379:methods*; D009446:Neurobiology; D059010:Single-Cell Analysis