Szöllösi J, Trón L, Damjanovich S, Helliwell S H, Arndt-Jovin D, Jovin T M
Cytometry. 1984 Mar;5(2):210-6. doi: 10.1002/cyto.990050216.
The energy transfer efficiency E was measured between fluorescein-conjugated concanavalin A (Con A) and rhodamine-conjugated Con A bound to homogeneous tissue culture cells, the HK22 murine lymphoma cell line. Results from a flow cytometric energy transfer method (FCET) and two different steady-state fluorimeter methods were compared. The data were found to be in close agreement after careful correction of the steady-state fluorimetric measurements for contributions from dissociating ligand. The biological variability of the individual cells with respect to E was calculated using an error propagation analysis and were found to be less than the variability in the absolute amount of ligand binding per cell. FCET has a number of advantages over the fluorimetric measurements using suspensions of cells: (1) relatively labile receptor-ligand complexes can be measured; (2) the analysis can be restricted to undamaged cells by gating the data collection on the light-scattering signals; (3) heterogeneous populations of cells with respect to donor and acceptor topology can be distinguished by the correlation of E with other cellular parameters derived from additional signals or combinations thereof; and (4) the dynamics of donor-acceptor redistribution on subpopulations can be measured.
在与均匀组织培养细胞HK22小鼠淋巴瘤细胞系结合的荧光素偶联伴刀豆球蛋白A（Con A）和罗丹明偶联Con A之间测量能量转移效率E。比较了流式细胞术能量转移方法（FCET）和两种不同的稳态荧光计方法的结果。在对解离配体的贡献进行仔细校正后，发现稳态荧光测量数据非常吻合。使用误差传播分析计算了单个细胞相对于E的生物学变异性，发现其小于每个细胞配体结合绝对量的变异性。与使用细胞悬液的荧光测量相比，FCET具有许多优点：（1）可以测量相对不稳定的受体-配体复合物；（2）通过根据光散射信号选通数据收集，可以将分析限制在未受损细胞上；（3）可以通过E与来自其他信号或其组合的其他细胞参数的相关性来区分供体和受体拓扑结构不同的细胞异质群体；（4）可以测量亚群上供体-受体重新分布的动力学。
