Originally called Mr. Zero, he was renamed and popularized by the 1960s Batman television series, in which he was played by several actors. Over two decades later, a television adaptation of Batman revitalized him once again. Batman: The Animated Series, retold Mr. Freeze's origin, introducing his terminally ill, cryogenically frozen wife, which greater explained his obsession with ice and need to build a criminal empire to raise research funds. The animated series version of Freeze was also more calm and composed than previous, campier versions and the comic book version has been altered to resemble it.

Elements of this personal tragedy, but not Freeze's calm demeanor, were incorporated into the 1997 film Batman & Robin, in which he was portrayed by Arnold Schwarzenegger.

From the time of his first appearance in 1959 onward, Mr. Freeze was portrayed as one of many "joke" villains (see also Killer Moth, The Mad Hatter) cast as stock enemies of Batman. Originally called Mr. Zero, the producers of the 1960s Batman television series renamed him Mr. Freeze (and portrayed Batman addressing him as "Dr. Schimmell"), and the name quickly carried over to the comic books.

Nearly thirty years later, Mr. Freeze would owe even more to television. In Batman: The Animated Series, the "Heart of Ice" episode, he was made into a more complex, tragic character. This version of Mr. Freeze was enthusiastically accepted by fans, and has become the standard portrayal for the character in most forms of media, including the comic book series itself, which previously had the character casually killed by the Joker. Freeze was hastily resurrected in the comic after the episode aired.

In the Pre-Crisis continuity, Mr. Freeze is a rogue scientist whose design for an "ice gun" backfires when he inadvertently spills cryogenic chemicals on himself, resulting in his needing sub-zero temperatures to survive.

In the Batman: Mr. Freeze special (based closely on the animated series episode "Heart of Ice") he is given a backstory. As a child, he is fascinated by freezing animals. His parents, horrified by his "hobby", send him to a strict boarding school, where he is miserable, feeling detached from humanity. In college, he meets a woman named Nora, whom he befriends and ultimately marries.

Nora later falls terminally ill. Fries takes on a job working for a large company run by the ruthless Ferris Boyle. Fries discovers a way to put Nora into cryo-stasis (using company equipment), hoping to sustain her until a cure could be found. Boyle finds out about the experiment and attempts to have her brought out of stasis, overruling Fries' frantic objections. A struggle ensues, in which Boyle kicks Fries into a table full of chemicals and leaves him for dead. Fries survives, but his body temperature is lowered dramatically; he can now only live at sub-zero temperatures, forced to wear a special refrigerating suit to stay alive. As Mr. Freeze, he uses cryonic technology to create a gun, which fires a beam that freezes any target within its range.

His first act as a costumed criminal is to take revenge upon Ferris Boyle, a plan with which Batman interferes. Mr. Freeze fires his freeze-gun at Batman, but he dodges, causing the beam to shatter Nora's capsule. Freeze blames Batman, and swears to destroy whatever the Dark Knight holds dear (mainly Gotham City, and eventually Robin).

Mr. Freeze's crimes tend to involve freezing everyone and everything he runs into. In addition, he hardly ever forges alliances with the other criminals in Gotham, preferring to work alone, although he has worked as a hired enforcer/hitman for the Black Mask. Once, in the hopes of reviving his wife, he allies himself with the Secret Society of Super Villains, fashioning for Nyssa al Ghul a sub-zero machine in exchange for the use of her own Lazarus Pit. He attempts to restore Nora to life without waiting for the adjusting needed in the pool chemicals, however; she returns to life as the twisted Lazara, and escapes. She blames her husband for her plight, and estranges herself from him.

He is usually imprisoned in Arkham Asylum when apprehended by Batman, as it is the only facility in Gotham that can accommodate his medical requirements for a refrigerated cell.

Most recently, he has been seen among the new Injustice League.

Like most Batman villains, Mr. Freeze plans his crimes about a specific theme; in his case, ice and cold. In darker incarnations of the Batman mythos, Mr. Freeze's obsession with ice stems from personal tragedy, and his crimes are inspired by his desire to make the rest of the world as cold and miserable as he is. He freezes areas around him using special weapons and equipment. His refrigeration suit grants him superhuman strength and durability, making him a powerful villain in Batman's rogues gallery. Some interpretations also suggest that because he has been soaked in the serum he intended to use for cryo-preservation, his age progression has slowed drastically. In the cartoon The Batman he has the power to generate ice and cold with his hands

In the No Man's Land storyline, Fries was even shown to be a formidable opponent for Superman.

In the Underworld Unleashed storyline, the demon Neron grants Mr. Freeze the ability to generate sub-zero temperatures, no longer needing his freeze-gun or refrigeration suit. Unlike most villains granted superpowers by the demon, Mr. Freeze's new abilities were only temporary.

According to Batman: The Animated Series, Freeze understands how to reproduce his powers. Upon being offered enough money to bankroll large scale research into Nora's condition, Freeze turns the wealthy, terminally ill Grant Walker into another "Mr. Freeze"-like being (on the theory that the "Mr. Freeze condition" would arrest the disease). However, Walker reveals that he wants to use his new abilities to turn the world into a frozen wasteland, leaving him and a few chosen followers to live eternally in Oceana, his underwater "World of Tomorrow". Freeze imprisons him in a block of ice Oceana collapses around him. For two years, Walker remains in the iceberg, driving him insane. Although he does not reappear in the animated series, the "second Mr. Freeze" reappeared in the comic book Batman: Gotham Adventures (based on the cartoon of the same name). In his last appearance, he breaks into the Wayne Foundation and kidnaps all of the scientists working to cure the original Mr Freeze. With Batman out of action due to a concussion, Batgirl, Nightwing, and Robin save the scientists, with the help of Mr. Freeze. Walker is eventually caught after a short fight with Freeze, and then sent to Arkham.