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.
| Original language | English (US) |
|---|---|
| Title of host publication | Three-Dimensional and Multidimensional Microscopy |
| Subtitle of host publication | Image Acquisition and Processing XXV |
| Editors | Tony Wilson, Carol J. Cogswell, Thomas G. Brown |
| Publisher | SPIE |
| Volume | 10499 |
| ISBN (Electronic) | 9781510614833 |
| DOIs | |
| State | Published - Jan 1 2018 |
| Event | Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV 2018 - San Francisco, United States Duration: Jan 29 2018 → Jan 31 2018 |
Publication series
| Name | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
|---|---|
| Volume | 10499 |
| ISSN (Print) | 1605-7422 |
Conference
| Conference | Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV 2018 |
|---|---|
| Country | United States |
| City | San Francisco |
| Period | 1/29/18 → 1/31/18 |
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Keywords
- Brain
- Fluorescence imaging
- Light-sheet microscopy
- Low-cost microscopy
ASJC Scopus subject areas
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Atomic and Molecular Physics, and Optics
- Radiology Nuclear Medicine and imaging
Cite this
}
Low cost light-sheet microscopy for whole brain imaging. / Kumar, Manish; Nasenbeny, Jordan; Kozorovitskiy, Yevgenia.
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV. ed. / Tony Wilson; Carol J. Cogswell; Thomas G. Brown. Vol. 10499 SPIE, 2018. 104991I (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 10499).Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
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
ER -