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. Author manuscript; available in PMC: 2015 Mar 1.

Published in final edited form as: Ann Biomed Eng. 2013 Nov 19;42(3):555–565. doi: 10.1007/s10439-013-0939-7

The 3D isotropic diffusion of FD500 molecule in glycerol/PBS solutions. (a) Typical time series of 3D MP-FRAP Z-stack image of fluorescent solutes for the experimental diffusion measurements in the glycerol/PBS solutions. For each experiment, typically, 40 stacks of post-bleaching images, plus 1 stack prior to photobleaching, are acquired at an approximate rate of 18 seconds/stack. The central region of the observation volume is photobleached by the multiphoton laser and fluorescence recovers completely in the last Z-stack image series (t=720s). (b) The results of 3D diffusion tensors (mean±SD). In each concentration group, there were no significant differences between the diagonal diffusion components (i.e., D_xx, D_yy, and D_zz), while the off-diagonal diffusion components were significantly smaller than the diagonal diffusion components (p<0.0001). Additionally, the nominal diffusivities (tr(D)/3) of FD500 significantly decreased (ANOVA, p<0.0001) with the increasing glycerol concentrations.

The 3D isotropic diffusion of FD500 molecule in glycerol/PBS solutions. (a) Typical time series of 3D MP-FRAP Z-stack image of fluorescent solutes for the experimental diffusion measurements in the glycerol/PBS solutions. For each experiment, typically, 40 stacks of post-bleaching images, plus 1 stack prior to photobleaching, are acquired at an approximate rate of 18 seconds/stack. The central region of the observation volume is photobleached by the multiphoton laser and fluorescence recovers completely in the last Z-stack image series (t=720s). (b) The results of 3D diffusion tensors (mean±SD). In each concentration group, there were no significant differences between the diagonal diffusion components (i.e., D_xx, D_yy, and D_zz), while the off-diagonal diffusion components were significantly smaller than the diagonal diffusion components (p<0.0001). Additionally, the nominal diffusivities (tr(D)/3) of FD500 significantly decreased (ANOVA, p<0.0001) with the increasing glycerol concentrations.