The BBC's Sport Relief TV appeal which included the live reunion of Kylie Minogue and Jason Donovan and a comedy appearance by David Beckham raised a record £51.2 million.
The fundraising spectacular beat the previous highest on-the-night total of £50.4 million.
The charity marathon on BBC One last night was hosted live for the first time from the Queen Elizabeth Olympic Park in east London and was kicked off by presenters Gary Lineker and David Walliams.
A host of stars featured in sketches to get the nation active and persuade them to donate to good causes that help thousands of people in the UK and across the world.
Lineker tweeted: "Over £51 million. Wonderful night and a treat to work with such brilliant people in front and behind the camera. Thanks."
Highlights included Beckham starring in the first new episode of Only Fools And Horses for over 11 years.
A bespectacled Beckham was in Peckham during the one-off special with Sir David Jason and Nicholas Lyndhurst. The retired England star poked fun at himself during a sketch in a greasy spoon cafe.
Australian singers Minogue and Donovan duetted on their 1988 smash hit Especially For You, which sold more than a million copies in the UK when it was first released.
The singers were joined on stage by comedian and co-presenter of the show Walliams whose aimed his affections at Donovan.
He jokingly tweeted: "Oh what a night!!! The lovemaking afterwards with @JDonOfficial was absolutely top-notch."
He later went on to present a special footballers edition of Celebrity Mastermind, with Tottenham defender Michael Dawson, Arsenal's Jack Wilshere, and Fulham midfielder, Scott Parker.
The Olympic-themed show also featured former gold medallist Lord Sebastian Coe taking part in a Clash of the Titans battle against comedian John Bishop at the London velodrome.
Lord Coe and Bishop captained teams in a four-event challenge, including a cycling elimination, synchronised swimming and rhythmic gymnastics.
The teams included a mix of sports stars and celebrities such as cricketer Andrew Flintoff, boxer Nicola Adams, singer Olly Murs and radio hosts Nick Grimshaw and Richard Bacon.
Flintoff won the men's cycling elimination for Team Bishop at the velodrome despite taking a tumble and being knocked off his bike.
Radio 1 DJ Greg James, who lost out to comedian Patrick Kielty in the synchronised swimming event, said afterwards: "WHAT A DAY! Loved it! HUGE total too for @sportrelief - well played everyone. I'm so so knackered."
Murs said: "Thanks for all your support tonight! @sportrelief was ace!! Raised a ton of money thanks to YOU lot!! Awesome. Sorry about the Lycra outfit!"
Britain's paralympians swapped their sports kits for sequins in a special version of Strictly Come Dancing.
Four athletes - blind footballer David Clarke, wheelchair racer Hannah Cockroft, former world champion javelin thrower Nathan Stephens and sitting volleyball player Martine Wright - danced to Katy Perry's Firework.
Wimbledon champion Andy Murray showed he was game for a laugh when comics on Mock The Week served up a volley of gags at his expense during a Sport Relief special.
The normally taciturn tennis star was a guest for an edition of the topical comedy show with panellists invited to give him a ''thrashing'', while his girlfriend Kim Sears looked on from the audience.
The show was kicked off by X Factor winners from 2011, Little Mix, performing the official Sport Relief single, Word Up, a cover of the 1986 hit by Cameo.
The band visited a hospital in Liberia where they saw children who are suffering from life threatening conditions from easily preventable diseases such as malaria.
Comedian Miranda Hart led the nation in an aerobics work-out accompanied by boxer David Haye and former footballer David Ginola, while Coldplay performed their single Magic.
The Government announced that it would cover the costs of the value-added tax on the Sport Relief single sales through funding from the Department for International Development, effectively giving back the VAT.
The biennial charity fund-raiser began in earnest before last night's live telethon with a number of celebrities having completed gruelling physical challenges to persuade people to donate.
On Thursday Radio 1 DJ Jo Whiley completed a 26-hour treadmill challenge, which ended with a £260,000 boost from supporters Coldplay.
Whiley had been in a perspex box outside Broadcasting House in London and was joined by a number of stars including celebrity cook Nigella Lawson.
In February, presenter Davina McCall accomplished a gruelling 500-mile cycle, swim and running journey from Edinburgh to London. McCall, who was rescued from freezing Lake Windermere, raised over £2.2 million from her challenge.
She said: "There were moments where it was amazing, really beautiful. Lake Windermere was my nemesis, but my hardest day was the first day bike ride. The wind coming in from Birmingham was the worst."
Whiley and McCall were taking part in the BBC Radio Around the World for Sport Relief, in which figures from BBC Radio are attempting to run, swim and cycle 25,000 miles which is roughly equivalent to the distance around the equator.
Co-host of The One Show Alex Jones was seen scaling 1,200ft up the mighty Moonlight Buttress in Zion National Park, Utah, in a three-day ordeal which raised over £1.2 million for her challenge.
She told hosts Lineker and Walliams: "I had bitten off more than I could chew because when we started in the first hour I realised what was in front of me and I felt so small. I'm not a climber and it was just so hard. It was petrifying for three days."
The show also included stark reminders of families in need across Britain. Appeals came from athlete Denise Lewis, actor Martin Shaw, presenter Emma Willis, model Abbey Clancy and singer Cheryl Cole. Half of all Sport Relief's donations go towards helping over 2,000 projects in the UK.
The show also included the first airing of the England football team's World Cup song, a cover version of Take That's Greatest Day.
Gary Barlow and former England captain Lineker led a 20-strong group including pop stars Katy B, former Spice Girl Emma Bunton, Pixie Lott and Conor Maynard.
Footballers Peter Shilton, Michael Owen, Glenn Hoddle, Dion Dublin and Sir Geoff Hurst also took part.
All proceeds from the sale of the record will go to Sport Relief.