TY - GEN
T1 - Low cost light-sheet microscopy for whole brain imaging
AU - Kumar, Manish
AU - Nasenbeny, Jordan
AU - Kozorovitskiy, Yevgenia
PY - 2018/1/1
Y1 - 2018/1/1
N2 - 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.
AB - 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.
KW - Brain
KW - Fluorescence imaging
KW - Light-sheet microscopy
KW - Low-cost microscopy
UR - http://www.scopus.com/inward/record.url?scp=85045341110&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85045341110&partnerID=8YFLogxK
U2 - 10.1117/12.2288497
DO - 10.1117/12.2288497
M3 - Conference contribution
AN - SCOPUS:85045341110
VL - 10499
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Three-Dimensional and Multidimensional Microscopy
A2 - Wilson, Tony
A2 - Cogswell, Carol J.
A2 - Brown, Thomas G.
PB - SPIE
T2 - Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV 2018
Y2 - 29 January 2018 through 31 January 2018
ER -