To keep the optimally physiological state of the strain in the process of glutamic acid fermentation, the one-factor-at-a-time method and response surface methodology experiment about the effect of glucose stress on the strains (FM00-187) in the fermentation medium were performed, and the physiological status (cell-viability) of corynebacterium glutamicum flora in different concentrations of glucose  was detected by Cyto Analysis 2000. With the changes in the glucose(substrate, initial glucose, fed-batch glucose) concentration, the cell-viability of the strains, changed significantly: increasing the concentration of the substrate, the cell-viability tended to increase .When the concentration of substrate glucose reached 8.93 g/L, the initial glucose concentration 39.14 g/L and the fed-batch glucose concentration 634.99 g/L, the cell-viability peaked (7.650µA), if the glucose continues to increase, the cell-viability would decrease, which was the phenomenon of glucose concentration inhibition. The sensitivity of cell-viability to the three factors, substrate glucose, fed-batch glucose ,initial glucose. Corynebacterium glutamicum fermentation performance can be displayed through the cell-activity, provides the reference for the actual fermentation production.