On Monday it was clear the problem that first flared up in Halle 11 days ago, when he had to quit midway through his semi-final, was again troubling him as he was stretched into a fifth set by Italy's Simone Bolelli.
He had the stricken left calf re-strapped while leading 3-0 in the decider and while that was enough to carry him over the finishing line in his first round match -- Monday's three and a half hour battle proved to be his undoing.
"It is same injury. Got better before the first match. I thought it was really going to be okay. But last match in fifth set I was hurting too much," said the dejected fifth seed who had been due to face Colombian Santiago Giraldo.
"I tried to warm-up today but it was hurting, you know, just walking and running, so I decided not to play today."
His exit will be a huge blow back in his homeland, where his every move is scrutinised by the country's 120 million plus population ever since he became the first Asian man to contest a grand slam singles final at the U.S. Open last year.
Following his victory over Bolelli, Nishikori had declared that the injury "should be okay for the next match" but it was a statement said in hope rather than with great belief.
"It's not a big injury, I think a little tear," added the Florida-based 25-year-old.
"During the first match against Simone, especially in the fifth set, it hurt pretty bad. I don't know how I fought through, but somehow I won.
"I knew it's going to be little pain I have next day... I thought it would get better in two days but it didn't.
"It's very disappointing. Especially as it's a grand slam. I thought I was playing well on grass. So very unfortunate to lose this chance," added Nishikori.
He now has a month off before returning to action in Washington for the American hard court swing.
(Reporting by Pritha Sarkar, editing by Justin Palmer and Martyn Herman)
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