Use of a membrane-bound fluorophore to characterize diffusion boundary layers around human erythrocytes.
Use of a membrane-bound fluorophore to characterize diffusion boundary layers around human erythrocytes.
复制标题
使用膜结合荧光团来表征人类红细胞周围的扩散边界层。
DOI：
10.1016/s0006-3495(86)83654-2
复制
发表时间：
1986
影响因子：
3.4
通讯作者：
Kutchai,H
中科院分区：
文献类型：
--
作者：
Williams,JB;Kutchai,H
一种新的方法被用来证明在停流分光光度计中快速混合后红细胞周围存在扩散边界层，并估计扩散边界层的表观尺寸。用四甲基罗丹明异硫氰酸酯(TRITC)标记的粉红色红细胞幽灵在停流装置中与50 mM NaI混合在林格氏溶液中。I-是TRITC荧光的有效碰撞猝灭剂。停流后TRITC荧光强度随时间呈单指数下降。在稳态实验中，根据TRITC荧光对I-浓度的依赖关系，估计了细胞表面I-浓度随时间的变化。细胞表面I-浓度增加的动力学符合两种扩散模型：由平面扩散边界层包围的平面红细胞幽灵和由球壳扩散边界层包围的球形红细胞。扩散边界层厚度为4.68微米时，平面模型与实验数据吻合较好。利用球面模型，用6.9微米扩散边界层对实验数据进行了最好的拟合。
A novel method is used to demonstrate the presence of diffusion boundary layers around erythrocytes following rapid mixing in a stopped-flow spectrophotometer and to estimate the apparent dimensions of the diffusion boundary layers. Pink erythrocyte ghosts labeled on their external surfaces with tetramethyl rhodamine isothiocyanate (TRITC) were mixed in a stopped-flow apparatus with 50 mM NaI in Ringer's solutions. I- is an effective collisional quencher of TRITC fluorescence. TRITC fluorescence after flow stopped decreased monoexponentially with time. The concentration of I- at the cell surface as a function of time was estimated from the dependence of TRITC fluorescence on I- concentration in steady-state experiments. The kinetics of the increase in I- concentration at the cell surface was fit to two diffusional models: a planar erythrocyte ghost bounded by planar diffusion boundary layer and a spherical erythrocyte surrounded by a spherical shell diffusion boundary layer. The planar model best fits the experimental data with a diffusion boundary layer 4.68 microns thick. Using the spherical model the experimental data is best fit by a 6.9 microns diffusion boundary layer.