The 64-year-old pop star took to the popular social media platform to come out of the closet.
She revealed her sexuality to her fans in a short, five-second-long clip. In the video, Madonna can be seen holding a pair of hot pink underwear.
The caption over the clip reads: "If I miss, I'm gay."
She then proceeds to throw the underwear in the direction of a bin, missing spectacularly.
She finishes the video with a shrug.
The social media clip took only moments to send the internet into overdrive, amassing over 1.3 million views in one hour.
Fans flooded the comments section, with many cheering on the 64-year-old star for seemingly confirming her sexuality, which has long been a topic of debate for music fans.
One user said: "What in the '80s is happening?"
Another added: "Did Madonna just come out? And I’m witnessing it in real time?"
A third chipped in with: "Are we like witnessing history in the making?"
Another just shared a shocked face emoji alongside one word: "WHAT?"
Others pointed out that Madonna has mentioned in the past she likes both men and women.
One user said: "Madonna has been an out bisexual for literally decades, my dudes."
Another added: "We already knew that."
Madonna has skirted around discussing her sexuality in the past, preferring not to fully admit her sexual preferences.
She has never confirmed or denied any of the rumours of dating women in the past, however The Sun states she has encouraged gossip about her sex life.
Back in 1990, shortly after her 'Justify My Love' video clip aired on television, the pop icon was open about her sexuality, as per the Daily Mail: "I think everybody has a bisexual nature. That’s my theory. I could be wrong."
18 years ago today, Britney Spears kissed Madonna at the VMAs in one of the most iconic performances in history pic.twitter.com/GxzPv26wge— Britney Stan 💕 (Joel Corry remix) (@BritneyTheStan) August 28, 2021
In a 1991 letter to supermodel Amanda Cazalet, which was made public by The Sun, said: "I think you are the most beautiful woman in the world. I’m dying to kiss you again. I fantasise about you all the time."
Madonna continued: "I cannot explain my attraction — nevertheless it’s there. I’m attracted to very few women in this way."
Madonna, Britney Spears, and Christina Aguilera also famously caused a stir on stage at the MTV VMA Awards with a steamy kiss that sent tongues wagging in 2003.