Rosol pulled off one of the biggest upsets in history when he knocked out the two-time champion on Centre Court, but lost 6-2 6-3 7-6 to Kohlschreiber.
The 26-year-old returned to form more akin to his ranking of 100, losing the first set in 25 minutes on Court 12.
Rosol saved one match point in the tie-break but lost in 96 minutes.
Meanwhile, Defending champion Petra Kvitova and four-time champion Serena Williams progresses to the fourth round.
Williams survived a scare to complete a 6-7 (5-7) 6-2 9-7 win over Jie Zheng while Kvitova eased through with a 6-1 6-0 victory over Varvara Lepchenko.