Zhang Yongliang, Chen Yu, Li David Day-Uei
Opt Express. 2016 Jun 27;24(13):13894-905. doi: 10.1364/OE.24.013894.
Fast deconvolution is an essential step to calibrate instrument responses in big fluorescence lifetime imaging microscopy (FLIM) image analysis. This paper examined a computationally effective least squares deconvolution method based on Laguerre expansion (LSD-LE), recently developed for clinical diagnosis applications, and proposed new criteria for selecting Laguerre basis functions (LBFs) without considering the mutual orthonormalities between LBFs. Compared with the previously reported LSD-LE, the improved LSD-LE allows to use a higher laser repetition rate, reducing the acquisition time per measurement. Moreover, we extended it, for the first time, to analyze bi-exponential fluorescence decays for more general FLIM-FRET applications. The proposed method was tested on both synthesized bi-exponential and realistic FLIM data for studying the endocytosis of gold nanorods in Hek293 cells. Compared with the previously reported constrained LSD-LE, it shows promising results.
快速去卷积是在大型荧光寿命成像显微镜（FLIM）图像分析中校准仪器响应的关键步骤。本文研究了一种基于拉盖尔展开的计算高效的最小二乘去卷积方法（LSD-LE），该方法最近被开发用于临床诊断应用，并提出了在不考虑拉盖尔基函数（LBFs）之间相互正交性的情况下选择LBFs的新标准。与先前报道的LSD-LE相比，改进后的LSD-LE允许使用更高的激光重复率，从而减少每次测量的采集时间。此外，我们首次将其扩展用于分析双指数荧光衰减，以用于更一般的FLIM-FRET应用。所提出的方法在合成的双指数和实际的FLIM数据上进行了测试，以研究Hek293细胞中金纳米棒的内吞作用。与先前报道的约束LSD-LE相比，它显示出了有前景的结果。
