一种毫微秒荧光谱去卷积处理方法及其在生物膜研究中的应用

DECONVOLUTION OF MACROMOLECLJLAR NANOSECOND FLUORESCENCE SPECTROSCOPY AND IT'S APPLICATION IN BIOMEMBRANE STUDIES

本文介绍了在大分子荧光寿命测量中,为消除测量系统响应时间的影响,对荧光谱进行去卷积处理的一种方法.该方法由实测的响应函数和荧光函数计算出响应矩阵和荧光矩阵,再用单指数函数拟合实测荧光谱的尾部,用迭代法求解.方法比较简单,也比较实用.我们用自制的NSY-1毫微秒荧光谱仪及所设计的微机程序,测试了荧光探剂Merocyanine 540(MC540)标记的脂质体以及重设线粒体H—ATP酶脂酶体的荧光寿命ι.它们的寿命分别为1.63ns和1.30ns.这一明显差别提示测量荧光寿命可作为研究生物膜膜脂—蛋白质相互作用的一种手段.

This paper introduces a method of deconvolution for analyzing nanosecond fluorescence spectroscopy of macromolecules, it can eliminate influence of instrumental response. The moments of instrumental response and sample fluorescence have been calculated by the aid of response function and fluorescence function measured at first, and then fit fluorescence curve at the tail to single exponential, the result is obtained by an iterative procedure. The moment method is reliabe and relatively simple.Fluorescence lifetimes of simple liposomes and proteoliposomes containing mitochondrial H+-ATPase labeled with Merocyanine 540 (MC540) have measured by NSY-1 nanoflumeter. The lifetime (τ) for former is 1.63ns and the later is 1.30ns. The difference is obvious. This may indicate that measurement of fluorescence lifetime of biomembrane samples with fluorescence probe is useful for studying interactions between membrane lipids and proteins.