Abstract
Two-photon microscopy is a very attractive tool for the study of the three-dimensional (3D) and dynamic processes in cells and tissues. One of the feasible constructions of two-photon microscopy is the combination a confocal laser scanning microscope and a mode-locked Ti:sapphire laser. Even though this approach is the simplest and fastest implementation, this system is highly cost-intensive and considerably difficult in modification. Many researcher therefore decide to build a more cost-effective and flexible system with a self-developed software for operation and data acquisition. We present a custom-built two-photon microscope based on a mode-locked Yb 3+ doped fiber laser and demonstrate two-photon fluorescence imaging of biological specimens. The mode-locked fiber laser at 1060 nm delivers 320 fs laser pulses at a frequency of 36 MHz up to average power of 80 mW. The excitation at 1060 nm can be more suitable in thick, turbid samples for 3D image construction as well as cell viability. The system can simply accomplish confocal and two-photon mode by an additional optical coupler that allows conventional laser source to transfer to the scanning head. The normal frame rate is 1 frames/s for 400 x 400 pixel images. The measured full width at half maximum resolutions were about 0.44 μm laterally and 1.34 μm axially. A multi-color stained convallaria, rat basophilic leukemia cells and a rat brain tissue were observed by two-photon fluorescence imaging in our system.
| Original language | English |
|---|---|
| Title of host publication | Three-Dimensional and Multidimensional Microscopy |
| Subtitle of host publication | Image Acquisition and Processing XIX |
| DOIs | |
| Publication status | Published - 2012 Apr 16 |
| Event | Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX - San Francisco, CA, United States Duration: 2012 Jan 24 → 2012 Jan 26 |
Publication series
| Name | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
|---|---|
| Volume | 8227 |
| ISSN (Print) | 1605-7422 |
Other
| Other | Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX |
|---|---|
| Country | United States |
| City | San Francisco, CA |
| Period | 12/1/24 → 12/1/26 |
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All Science Journal Classification (ASJC) codes
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Atomic and Molecular Physics, and Optics
- Radiology Nuclear Medicine and imaging
Cite this
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A custom-built two-photon microscope based on a mode-locked YbZ 3+ doped fiber laser. / Kim, Dong Uk; Song, Hoseong; Song, Woosub; Kwon, Hyuk Sang; Kim, Dug Yong.
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX. 2012. 82271F (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 8227).Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
TY - GEN
T1 - A custom-built two-photon microscope based on a mode-locked YbZ 3+ doped fiber laser
AU - Kim, Dong Uk
AU - Song, Hoseong
AU - Song, Woosub
AU - Kwon, Hyuk Sang
AU - Kim, Dug Yong
PY - 2012/4/16
Y1 - 2012/4/16
N2 - Two-photon microscopy is a very attractive tool for the study of the three-dimensional (3D) and dynamic processes in cells and tissues. One of the feasible constructions of two-photon microscopy is the combination a confocal laser scanning microscope and a mode-locked Ti:sapphire laser. Even though this approach is the simplest and fastest implementation, this system is highly cost-intensive and considerably difficult in modification. Many researcher therefore decide to build a more cost-effective and flexible system with a self-developed software for operation and data acquisition. We present a custom-built two-photon microscope based on a mode-locked Yb 3+ doped fiber laser and demonstrate two-photon fluorescence imaging of biological specimens. The mode-locked fiber laser at 1060 nm delivers 320 fs laser pulses at a frequency of 36 MHz up to average power of 80 mW. The excitation at 1060 nm can be more suitable in thick, turbid samples for 3D image construction as well as cell viability. The system can simply accomplish confocal and two-photon mode by an additional optical coupler that allows conventional laser source to transfer to the scanning head. The normal frame rate is 1 frames/s for 400 x 400 pixel images. The measured full width at half maximum resolutions were about 0.44 μm laterally and 1.34 μm axially. A multi-color stained convallaria, rat basophilic leukemia cells and a rat brain tissue were observed by two-photon fluorescence imaging in our system.
AB - Two-photon microscopy is a very attractive tool for the study of the three-dimensional (3D) and dynamic processes in cells and tissues. One of the feasible constructions of two-photon microscopy is the combination a confocal laser scanning microscope and a mode-locked Ti:sapphire laser. Even though this approach is the simplest and fastest implementation, this system is highly cost-intensive and considerably difficult in modification. Many researcher therefore decide to build a more cost-effective and flexible system with a self-developed software for operation and data acquisition. We present a custom-built two-photon microscope based on a mode-locked Yb 3+ doped fiber laser and demonstrate two-photon fluorescence imaging of biological specimens. The mode-locked fiber laser at 1060 nm delivers 320 fs laser pulses at a frequency of 36 MHz up to average power of 80 mW. The excitation at 1060 nm can be more suitable in thick, turbid samples for 3D image construction as well as cell viability. The system can simply accomplish confocal and two-photon mode by an additional optical coupler that allows conventional laser source to transfer to the scanning head. The normal frame rate is 1 frames/s for 400 x 400 pixel images. The measured full width at half maximum resolutions were about 0.44 μm laterally and 1.34 μm axially. A multi-color stained convallaria, rat basophilic leukemia cells and a rat brain tissue were observed by two-photon fluorescence imaging in our system.
UR - http://www.scopus.com/inward/record.url?scp=84859588744&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84859588744&partnerID=8YFLogxK
U2 - 10.1117/12.907648
DO - 10.1117/12.907648
M3 - Conference contribution
AN - SCOPUS:84859588744
SN - 9780819488701
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Three-Dimensional and Multidimensional Microscopy
ER -