A custom-built two-photon microscope based on a mode-locked YbZ 3+ doped fiber laser

Dong Uk Kim, Hoseong Song, Woosub Song, Hyuk Sang Kwon, Dug Yong Kim

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Citations (Scopus)


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 languageEnglish
Title of host publicationThree-Dimensional and Multidimensional Microscopy
Subtitle of host publicationImage Acquisition and Processing XIX
Publication statusPublished - 2012
EventThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX - San Francisco, CA, United States
Duration: 2012 Jan 242012 Jan 26

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ISSN (Print)1605-7422


OtherThree-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX
Country/TerritoryUnited States
CitySan Francisco, CA

Bibliographical note

Funding Information:
Higher Education for its MyBrain15scholarship to pursue PhD programme in Halal Products Research Institute, Universiti Putra Malaysia (UPM). This research project was supported by Universiti Putra Malaysia (UPM) for providing the funding support awarded to Prof. Dr. Russly Ab Rahman through Project No. 9316900.

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Biomaterials


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