23 February 2018 Low cost light-sheet microscopy for whole brain imaging
Author Affiliations +
Abstract
Light-sheet microscopy has evolved as an indispensable tool in imaging biological samples. It can image 3D samples at fast speed, with high-resolution optical sectioning, and with reduced photobleaching effects. These properties make light-sheet microscopy ideal for imaging fluorophores in a variety of biological samples and organisms, e.g. zebrafish, drosophila, cleared mouse brains, etc. While most commercial turnkey light-sheet systems are expensive, the existing lower cost implementations, e.g. OpenSPIM, are focused on achieving high-resolution imaging of small samples or organisms like zebrafish. In this work, we substantially reduce the cost of light-sheet microscope system while targeting to image much larger samples, i.e. cleared mouse brains, at single-cell resolution. The expensive components of a lightsheet system – excitation laser, water-immersion objectives, and translation stage – are replaced with an incoherent laser diode, dry objectives, and a custom-built Arduino-controlled translation stage. A low-cost CUBIC protocol is used to clear fixed mouse brain samples. The open-source platforms of μManager and Fiji support image acquisition, processing, and visualization. Our system can easily be extended to multi-color light-sheet microscopy.
© (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Manish Kumar, Manish Kumar, Jordan Nasenbeny , Jordan Nasenbeny , Yevgenia Kozorovitskiy, Yevgenia Kozorovitskiy, } "Low cost light-sheet microscopy for whole brain imaging", Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 104991I (23 February 2018); doi: 10.1117/12.2288497; https://doi.org/10.1117/12.2288497
PROCEEDINGS
6 PAGES


SHARE
Back to Top