Queen Elizabeth's son Prince Andrew, her grandson Harry and his wife Meghan will not join her on the Buckingham Palace balcony for this year's Trooping the Colour, royal officials said on Friday.
Instead, the 96-year-old monarch has decided to limit numbers for the traditional set-piece appearance on her official birthday celebration to working royals only.
"Only those members of the royal family who are currently undertaking official public duties on behalf of the queen" will join her on June 2, a spokesman said.
The decision was taken "after careful consideration", he added.
Speculation had mounted that all three could be at the event, which kicks off four days of public celebrations for the queen's Platinum Jubilee.
Andrew, 62, in March made his first public appearance since settling a US civil claim for sexual assault, and after public outrage at his friendship with convicted sex offender Jeffrey Epstein.
The former Royal Navy helicopter pilot provided his mother, who has been in ill health and had difficulty walking and standing, with a steadying arm at a memorial service to her late husband, Prince Philip.
His prominent role at the televised Westminster Abbey event was seen as a sign that his mother believed he still has a part to play at family occasions.
But his appearance caused controversy and dominated coverage of the memorial service and Friday's announcement may be seen as a sign the palace does not want a repeat.
Andrew has strenuously denied the assault claims and remains stripped of his honorary military titles and charities, giving him no official royal role.
Senior royals
Speculation that Harry would return from his self-imposed exile in California has also increased after he visited his grandmother at her Windsor Castle home last month.
The 37-year-old former British Army captain is the second son of her eldest son and heir Prince Charles, and his first wife, Princess Diana.
He quit royal life in March last year, moving to the United States with his wife Meghan, where both have publicly complained about life in Britain's most famous family.
The couple, who still use their official titles of the Duke and Duchess of Sussex, are now involved in charitable and philanthropic work.
Denying them a place on the balcony will likely be seen as the queen refusing to accept a "half-in, half-out" approach to royal duties.
There was no immediate word on whether all three would be involved in the other public events to mark the queen's record-breaking 70 years on the throne.
But a spokeswoman for Harry and Meghan indicated they would be there with their children Archie, who turned three on Friday, and Lilibet, who was born in June last year and has yet to meet her great-grandmother.
The couple was "excited and honoured to attend the queen's Platinum Jubilee celebrations this June with their children", she added.
The decision means the queen will be joined on the balcony for the end of the military pageant and ceremonial fly-past by senior royals led by Charles and his second wife, Camilla.
Harry's older brother Prince William and his wife Kate will be there, with their children, Prince George, Princess Charlotte and Prince Louis.
The queen's youngest son, Prince Edward, will be accompanied by his wife, Sophie, and their children Louise and James.
Her only daughter, Princess Anne, will also be on the balcony. The queen has allowed her second husband, Tim Laurence, even though he is not a working royal.
The palace spokesman said the queen recognised him as a "frequent attendee and support for the Princess Royal (Anne) on official engagements".