Three new six-branched chromophores with different isolation groups were designed and synthesized by incorporating azo chromophores onto phenyl glycerol ester via an esterification reaction. The chemical structures were confirmed by 1H NMR, 13C NMR, FT-IR spectroscopy, mass spectrometry, element analysis and UV-visible absorption spectra. The six-branched dendritic chromophores can form thin films directly without adding any polymer matrix and show a quite high chromophore loading density. After electric poling, the nonlinear optical coefficients (d33) of the dendrimer films were determined to be around 113, 142 and 136 pm V−1, which are almost 3 times the maximum d33 value of the azo-chromophore doped polymer films. The further improvement in optical nonlinearity was successfully realized by mixing the six-branched chromophores containing different isolation groups.