Samurai (侍) are the guardians and military power of the Land of Iron. Little is known about them, although Ōnoki indicates that any shinobi would be foolish enough to challenge the military of the Land of Iron. Their general is Mifune. The Land of Iron is the only power in the modern world which continues to use samurai, as Hanzō alluded to that in the past, even in his time they were being used less and less, being replaced by ninja, ninjutsu, and ninja villages.
They are usually seen adorned in heavy plate-armor along with helmets and face masks that act like respirators, designed intentionally to combat poison regularly used by shinobi, and which include inbuilt radios, with senior officers having short horns atop of their helmets to identify themselves. Like real samurai, they are armed with swords but the scabbards of these, are instead mounted directly onto their faulds. However, the number of blades worn by a single warrior is also unusual, going as high as four, in spite of the fact that they have never been observed using more than two at a time in battle, which could probably mean they are kept as spares.
Little is known about the abilities of the samurai even by shinobi, but they are primarily sword-users. It was shown that they are capable of utilising chakra, apparently by channelling it through their swords. This allows them to extend the reach of their swords as well as attack at a distance by swinging them and releasing crescents of energy at their targets. The samurai have also shown form manipulation, as the chakra blades they form into many types, such as buzz-saws, broad swords, and axes. One of the forms of kenjutsu they use is Iaidō (居合道) which, due to the extremely quick drawing and re-sheathing of the katana, makes ninjutsu near useless since the hand seals might not be completed. As Ōnoki had stated that anyone daring to challenge the Land of Iron's military power would be considered foolish, this supports the formidable force of the samurai.
- Earlier in the series, Gatō had two samurai bodyguards, Zōri and Waraji, in his employ. Whether they were originally from the Land of Iron, or were taught by another organisation, is unknown.
- In the first movie, Sandayū Asama led a group of samurai to attack Dotō Kazahana.
- Samurai are traditionally known to use the katana, but the standard-issue sword wielded by the samurai of the Land of Iron seem to be closer in size and appearance to wakizashi. It is possible that samurai who can use chakra don't need a full-sized katana, as their Samurai Sabre Technique more than compensates for the reach that a wakizashi lacks.
- Curiously, though traditionally ninja weapons, in the weapons arsenal of the samurai of the Land of Iron, kusarigama were seen hanging on a wall.