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首页> 外文期刊>Microscopy research and technique >Differences between FLIM phasor analyses for data collected with the Becker and Hickl SPC830 card and with the FLIMbox card
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Differences between FLIM phasor analyses for data collected with the Becker and Hickl SPC830 card and with the FLIMbox card

机译：FLIM phasor分析与Becker和Hickl SPC830卡收集的数据与Flimbox卡进行分析的差异

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摘要

The phasor approach to FLIM (Fluorescence Lifetime Imaging Microscopy) is becoming popular due to the powerful fit free analysis and the visualization of the decay at each point in images of cells and tissues. However, although several implementation of the method are offered by manufactures of FLIM accessories for microscopes, the details of the conversion of the decay to phasors at each point in an image requires some consideration. Here, we show that if the decay is not properly acquired, the apparently simple phasor transformation can provide incorrect phasor plots and the results may be misinterpreted. In particular, we show the disagreement in experimental data acquired on the same samples using the two cards (FLIMbox, frequency domain and Becker & Hickl BH 830, time domain) and the effect produced by using the BH 830 card with different settings. This difference in data acquisition translates to the assignment of phasor components calculated using different acquisition parameters. This effect is already present in the original data that are not acquired with the proper parameters for the phasor conversion. We also show that the difference in the resolution of components already exists in the data acquired in the time domain when used with settings that do not allow acquisition of the fluorescence decay on a sufficient large time scale.
机译：由于细胞和组织图像图像和组织的图像中的每个点处的衰减的强大的自由分析和衰减的可视化，相移方法（荧光寿命显微镜显微镜显微镜）正在变得流行。然而，尽管通过用于显微镜的Flim附件制造商提供了几种方法，但是在图像中每个点处的衰减转换到相量的细节需要一些考虑。在这里，我们表明，如果没有正确获得衰变，显然简单的量量转换可以提供不正确的量程图，结果可能会误解。特别是，我们展示了使用两张卡（Flimbox，频域和Becker和Hickl BH 830，时域）在相同样本上获取的实验数据中的分歧，并通过使用不同的设置使用BH 830卡产生的效果。数据采集​​中的这种差异转化为使用不同采集参数计算的相分散组件的分配。该效果已经存在于未在Phasor转换的适当参数获取的原始数据中。我们还表明，当与不允许在足够大的时间尺度上采集的设置时，在时域中获取的数据中已经存在的分辨率的分辨率差异已经存在于时域中的数据。

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