대한기계학회논문집B (Transactions of the Korean Society of Mechanical Engineers B)

  • 제36권5호
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  • Pages.533-538
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  • 2012
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  • 1226-4881(pISSN)
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  • 2288-5324(eISSN)
대한기계학회 (The Korean Society of Mechanical Engineers)
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산소 민감 발광 염료를 이용한 마이크로 채널 내에서 배양되는 세포 주변의 산소 농도 측정

Method for the Measurement of Dissolved Oxygen in a Cell Culture Microchannel Using Oxygen-Sensitive Luminescence

  • 이승열 (한국산업기술대학교 지식기반에너지 대학원) ;
  • 진송완 (한국산업기술대학교 기계공학과)
  • Lee, Seung-Youl (Dept. of Mechanical System Engineering,Graduate School of Knowledge-Based Technology and Energy, Korea Polytechnic Univ.) ;
  • Jin, Song-Wan (Dept. of Mechanical System Engineering, Korea Polytechnic Univ.)
  • 투고 : 2011.12.30
  • 심사 : 2012.02.20
  • 발행 : 2012.05.01
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초록

본 논문에서는 산소 민감 발광 염료를 이용하여 마이크로 채널 내에서 세포가 배양되고 있을 때 산소농도를 측정하였다. 현재까지 알려진 여러 산소 민감 발광 염료 중 본 논문에서는 물에 잘 녹으며 장 시간 동안 사용하여도 독성이 없는 것으로 알려진 $[Ru(bpy)_3]^{2+}$를 사용하였으며 이와 더불어 산소 민감 염료 측정법의 단점을 보완하기 위하여 칼세인 염료를 이용하여 두 염료의 밝기 비율을 구하여 농도를 측정하였다. SCOMS 카메라와 마이크로 채널을 이용하여 캘리브레이션을 실시하고 농도와 밝기와의 관계를 구하였으며 이 관계를 이용하여 세포가 배양되는 조건에서의 배양액의 산소농도를 측정하였다. 실험 결과 채널입구에서 점점 멀어질수록 마이크로 채널 내에서 산소농도는 점점 낮아진다는 것을 관찰 할 수 있었다.

In this study, we used an $O_2$-sensitive luminescent dye to measure the $O_2$ concentration of culture media around HeLa cells cultured in a microchannel. $[Ru(bpy)_3]^{2+}$, which dissolves easily in water and which has no phototoxic effect, was used as the $O_2$-sensitive dye. The ratiometric sensing method was applied by introducing calcein as the $O_2$-insensitive dye, in order to overcome the disadvantages of intensity-based sensing. By performing calibration with an amperometric $O_2$ sensor, we could calculate the exact concentration of $O_2$ in the culture media. We applied this technique to measure the $O_2$ concentration around the cultured cells in the microchannel. As expected, the $O_2$ concentration gradually decreased as the cells moved farther away from the channel. This method is expected to be applicable to the investigation of hypoxia, which occurs commonly in scaffolds.

키워드

참고문헌

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피인용 문헌

  1. Ratiometric Dissolved Oxygen Sensors Based on Ruthenium Complex Doped with Silver Nanoparticles vol.17, pp.3, 2017, https://doi.org/10.3390/s17030548