Department of Chemical Engineering, University of Delaware, Newark, Delaware 19716, USA.
Biotechnol Bioeng. 1993 Nov 20;42(10):1190-8. doi: 10.1002/bit.260421009.
Bioluminescence has recently become a popular research tool in several fields, including medicine, pharmacology, biochemistry, bioprocessing, and environmental engineering. Beginning with purely qualitative goals, scientists are now targeting more demanding applications where accurate, quantitative interpretation of bioluminescence is necessary. Using the recent advances in fiber-optic technology, bioluminescence is easily monitored in vivo and in real time. However, the convenience of this measurement is often concealing an unsuspected problem: the bioluminescence signal might be corrupted by a large error caused by the extinction of light by biological cells. Since bioluminescent cultures not only emit light but also absorb and scatter it, the measured signal is related in a complex, nonlinear, and cell-concentration-dependent manner to the "true" bioluminescence. This light extinction effect, known as the "inner filter effect," is significant in high-density cultures. Adequate interpretation of the bioluminescence signal can be difficult without its correction. Here, we propose a real-time algorithm for elimination of the inner filter effect in a bioreactor. The algorithm yields the bioluminescence which would be measured if the glowing culture was completely transparent. This technique has been successfully applied to batch and continuous cultivation of recombinant bioluminescent Escherichia coli.
生物发光最近已成为多个领域（包括医学、药理学、生物化学、生物加工和环境工程）的热门研究工具。最初的目标是纯定性的，但现在科学家们的目标是更具挑战性的应用，这些应用需要对生物发光进行准确、定量的解释。利用光纤技术的最新进展，生物发光可以轻松地在体内实时监测。然而，这种测量的便利性常常掩盖了一个意想不到的问题：生物发光信号可能会因生物细胞的光损耗而导致大误差，从而受到严重干扰。由于发光培养物不仅会发射光，还会吸收和散射光，因此测量的信号与“真实”生物发光之间存在复杂的、非线性的、且与细胞浓度相关的关系。这种光损耗效应被称为“内滤效应”，在高密度培养物中非常显著。如果不进行校正，对生物发光信号进行充分解释可能会很困难。在这里，我们提出了一种用于消除生物反应器中内滤效应的实时算法。该算法可以得出如果发光培养物完全透明时将测量到的生物发光。该技术已成功应用于重组生物发光大肠杆菌的分批和连续培养。
