The world No 122 Broady insisted Ostapenko should have been disqualified after the Latvian threw her racquet and it bounced off the ground and hit a ballboy in their second set tiebreak.
The ball boy was called to the umpires' chair, and confirmed the racquet had hit him on the shoulder. As emotions ran high, Broady, who was trailing by a set at the time, was in tears as she first remonstrated with umpire Blaze Trifunobski, then tournament supervisor Tony Cho.
Ostapenko was just as incredulous, claiming that the incident was an accident. Ostapenko clearly threw her racquet, though it didn't appear to be directed intentionally at the ballboy.
"It was quite clear that she threw her racquet and it hit a ball kid...which should be [instant] disqualification," said Broady. "Jelena said to [the umpire] it slipped from her hand so it was just a code violation or something. On the replay it is quite clear but that's tennis and I managed to keep myself together quite well."
The WTA rules state that players can be defaulted for "unsporting" actions on court. Tim Henman was disqualified from a doubles match in his first year at Wimbledon in 1995, when he hit a ball in frustration at the end of a point and it struck a ball girl in the head.
"The ball kids are volunteers and a lot of them are quite young so you need to have a lot of respect for them," added Broady. "I don't think it was good behaviour."
There was obvious animosity between the players for the rest of the match, and continued at the end of the game, which was eventually won by Broady 4-6 7-6 (4) 7-5. After a cursory handshake, the duo continued to snipe at each other before the umpire stepped in and Ostapenko left the court.
"At the end of the match Jelena commented that my behaviour was terrible, which I thought was a bit out of order," said Broady. "Tensions were high, adrenalin was going so we had a bit of a confrontation."
In other action, two-time grand slam champion Svetlana Kuznetsova never looked comfortable against Tamira Paszek and was dismissed 6-2 6-3 in just over 90 minutes.
Another seed had fallen, but Paszek is becoming one of the stories of the tournament, after progressing through three matches in qualifying and beating Francesca Schiavone in a three hour first round marathon. Fellow qualifier Kirsten Flipkens also progressed, beating American Irina Falconi 6-1 4-6 6-2.
Fifth seed Sloane Stephens brought some normality to proceedings, coming back from a set down to beat German world No 64 Carina Witthoeft 3-6 6-1 6-3. Stephens was broken twice in the opening set but almost impeccable after that, only dropping one more service game across the rest of the match.
Russian teenager Daria Kasatkina couldn't repeat her heroics of Tuesday, when she took down Venus Williams, losing 5-7 6-2 5-7 to Japan's world No 66 Nao Hibino. Czech veteran Barbora Strycova was the sixth seeded player in the tournament to depart, losing 6-4 7-6 (7) to Julia Goerges.