The crew were called to a horse-riding accident in the Clyde Valley on Sunday, marking the milestone eight months after its first flight.
Scotland's Charity Air Ambulance (SCAA) is based at Perth Airport and supplements the work of the Scottish Ambulance Service across the country.
It is crewed by paramedics and operates 10 hours a day, seven days a week.
It costs about £1.5 million each year, made up from public and private donations and contributions from other charities.
The helicopter has flown more than 17,800 miles since May last year. SCAA chief executive Gavin Davey said: "Our crews have dealt with all types of emergency from road traffic accidents and serious falls to medical emergencies such as heart attacks and strokes.
"The people of Scotland seem to have taken SCAA to their heart and they know that the only fuel we fly on is their charity giving.
"Our growing band of supporters continues to step up to the mark and ensure Helimed 76 keeps flying for the benefit of everyone in the country.
"There is no doubt that this unique service in Scotland has contributed to saving numerous lives and relieving pain, and SCAA's work is impacting on the lives of hundreds of people right across Scotland - either directly with patients or indirectly with relatives, friends and colleagues."
Shortly after its launch the operating hours of the helicopter were changed to meet demand.
It started working between 8am and 6pm but the pattern of call-outs showed that the vast majority of deployments happen later in the day, so new operational times of 10am to 8pm were introduced.