The Arsenal midfielder had missed the national side's two warm-up games with a calf setback, but has been given the all-clear to play against Russia by head coach Michal Bilek
The creative midfielder had missed both of his side's warm-up games against Israel and Hungary after sustaining a calf injury in Arsenal's last Premier League match of the season against West Brom.
But despite a replacement being called up as a precaution, he has now been declared fit to participate in the Czechs' European campaign, which kicks off against Dick Advocaat's men.
"Rosicky will play in the opening match," Bilek told reporters. "He showed his qualities even during training.
"When he gets the ball, he is able to find space for a clever pass. He is decisive with the ball.
"Today showed how much he is looking forward to playing. I believe that he will boost the quality of our lineup."
Czech Republic will also face Greece and co-hosts Poland on the June 12 and June 16 respectively as they look to reach the quarter-finals of the competition.