Plastic damage assessment in 316 austenitic steel was performed in this research by using the misorientation parameters derived from an in situ EBSD technique. With the increase in plastic strain, the misorientation parameters, such as the Grain Reference Orientation Deviation (GROD), Grain Orientation Spread (GOS), the Grain Orientation Spread over the grain Diameter (GOS/D), and Geometrically Necessary Dislocation (GND) density presented a growing trend. Nevertheless, the variation in GROD did not show a monotony trend, and the relative increase in the amplitude of GOS and GND density was less in the late plastic stage. Compared with the above parameters, the (GOS)/D exhibited a near-linear increase during the plastic tensile stage. As the specimen was stretched to a strain of 56.99%, the (GOS)/D increased 8.9 times compared with the original specimen. The results showed that the (GOS)/D parameter has the potential of becoming an indicator for the assessment of plastic damage in 316 steel.