Petrášek Zdeněk, Bolivar Juan M, Nidetzky Bernd
Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, NAWI Graz, Graz, Austria.
Austrian Centre of Industrial Biotechnology, Graz, Austria.
Microsc Res Tech. 2021 Jan;84(1):71-78. doi: 10.1002/jemt.23566. Epub 2020 Jul 28.
We determine the optimal parameters (scan velocities) for measuring the luminescence lifetime on the microsecond scale using the recently introduced method based on scanning the excitation beam across the sample. Using simulations, we evaluate the standard deviation and bias of the luminescence decay rate determined by scanning with two different velocities. The analysis is performed for Poisson- and normal-distributed signals, representing different types of detection techniques. We also show that a weak uncorrected background induces a bias in the obtained decay rate, and take this effect into account when choosing optimal measurement parameters. For comparison, the analysis is additionally performed for two conventional gating schemes for lifetime measurement. The variable-velocity scanning method is found to be more robust to the effect of the background signal than the gating schemes.
我们使用最近引入的基于在样品上扫描激发光束的方法,确定了用于在微秒尺度上测量发光寿命的最佳参数(扫描速度)。通过模拟,我们评估了以两种不同速度扫描所确定的发光衰减率的标准偏差和偏差。针对泊松分布和正态分布信号进行了分析,它们代表不同类型的检测技术。我们还表明,微弱的未校正背景会在获得的衰减率中引起偏差,并在选择最佳测量参数时考虑到这种影响。为作比较,还针对两种用于寿命测量的传统选通方案进行了分析。发现变速扫描方法比选通方案对背景信号的影响更具鲁棒性。