Journal of the Korean Society of Visualization (한국가시화정보학회지)

The Korean Society of Visualization (한국가시화정보학회)
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Development of Image-based Fluorescence Photobleaching Technique for Measuring Macromolecule Diffusion in Biological Porous Medium

생체 다공성 매질에서 분자 확산 측정을 위한 영상 기반 형광 광표백 기법 개발

  • 이동희 (국민대학교 대학원 기계공학과) ;
  • 이정훈 (서울대학교 정밀기계설계공동연구소) ;
  • 박춘호 (국민대학교 기계자동차공학부) ;
  • 김중경 (국민대학교 기계자동차공학부)
  • Published : 2009.08.31
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Abstract

Fluorescence recovery after photobleaching (FRAP) has been widely used for the measurement of molecular diffusion in living cells and tissues. We developed an image-based FRAP (iFRAP) technique using a modified real-time microscope and a 488 nm Ar-ion laser. A fractional intensity curve was obtained from the time-lapse images of fluorescence recovery in the bleached spot to determine the diffusion coefficient of fluorescently labeled macromolecules in porous medium. We validated iFRAP through experiments with agar gels (0.5% and 1.5% w/v) containing FITC-Dextrans (10, 70 and 500 kDa MW). Further validation was performed by a Monte Carlo approach, where we simulated the three-dimensional random walk of macromolecules in agar gel model. Diffusion coefficients were deduced from the mean square displacement curves and showed good agreements with those measured by iFRAP.

Keywords

References

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