Aiming for promoting cell-immobilization in the bioreactor and enhancing continuous hydrogen production, a novel groove-type flat panel photo-biological reactor (GFPR) was developed. Photosynthetic bacteria (PSB) strain of Rhodopseudomonaspalustris was successfully attached on the groove-type surface of transparent material and generated PSB type biofilm. Strategies on improving continuous photo-biological hydrogen production within GFPR were comprehensively investigated. Experimental results revealed that hydrogen production rate, substrate degradation efficiency and light conversion efficiency of GFPR were obviously increased to 1.17 mmol/(L ·h),77.5% and 20.15%, respectively, under specific operating conditions of 590 nm of light wavelength, 9 W/m 2 of light intensity, 55 mmol/L of inlet substrate concentration and 960 mL/h of flow rate with glucose-based medium. Methods such as choosing proper light wavelength, light intensity to accelerate photophosphorylation for photosynthetic bacteria to accomplish hydrogen production metabolism and enhancing substrate transportation using convective mass transfer process were proved to be the effective way for promoting performance of photo-biological hydrogen production within the cell immobilized reactor operated under continuous flow mode. The experiment results were introduced to further research photo-biological reactor for practical hydrogen production.