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Development of line-scanning two-photon microscopy based on spatial and temporal focusing for tryptophan based auto fluorescence imaging

고속 트립토판 자가형광 이미징을 위한 시공간적 집중 기반의 라인 스캐닝 이광자 현미경 개발

  • 이준호 (포항공과대학교 기계공학과) ;
  • 남효석 (포항공과대학교 기계공학과) ;
  • 김기현 (포항공과대학교 기계공학과)
  • Received : 2013.09.05
  • Accepted : 2013.09.26
  • Published : 2013.09.30

Abstract

Two-photon microscopy (TPM) is minimally-invasive 3D fluorescence microscopy based on nonlinear excitation, and TPM can visualize cellular structures based on auto-fluorescence. Line-scanning TPM is one of high-speed TPM methods without sacrificing the image resolution by using spatial and temporal focusing. In this paper, we developed line-scanning TPM based on spatial and temporal focusing for auto-fluorescence imaging by exciting the tryptophan. Laser source for this system was an optical parametric oscillator (OPO) and it made near 570 nm femtosecond pulse laser. It had 200fs pulse width and 1.72 nm bandwidth, so that the achievable depth resolution was 2.41um and field of view (FOV) is 10.8um. From the characterization, our system has 3.0 um depth resolution and 12.3 um FOV. We visualized fixed leukocyte cell sample and compared with point scanning system.

Acknowledgement

Supported by : 한국과학재단

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