Bio-actuators that use insect muscular tissue have attracted attention from researchers worldwide because of their small size, self-motive property, self-repairer ability, robustness, and the need for less environment management than mammalian cells. To demonstrate the potential of insect muscular tissue for use as bio-actuators, three types of these robots, a pillar actuator, a walker, and a twizzer, have been designed and fabricated. However, a model of an insect muscular tissue-powered swimming robot that is able to float and swim in a solution has not yet been reported. Therefore, in this paper, we present a prototype of an insect muscular tissue-powered autonomous micro swimming robot that operates at room temperature and requires no temperature and pH maintenance. To design a practical robot body that is capable of swimming by using the force of the insect dorsal vessel (DV), we first measured the contraction force of the DV. Then, the body of the swimming robot was designed, and the design was confirmed by a simulation that used the condition of measured contraction force. After that, we fabricated the robot body using polydimethylpolysiloxane (PDMS). The PDMS body was obtained from a mold that was fabricated by a stereo lithography method. Finally, we carefully attached the DV to the PDMS body to complete the assembly of the swimming robot. As a result, we confirmed the micro swimming robot swam autonomously at an average velocity of 11.7 µm/s using spontaneous contractions of the complete insect DV tissue. These results demonstrated that the insect DV has potential for use as a bio-actuator for floating and swimming in solution.2 of 15 considerable attention, and bio-hybrid microdevices (the body structure can be made from polymer) were reported, including a pillar actuator and micro heart pumps [7][8][9][10][11], and different kinds of robots, including walking and swimming robots [12][13][14][15][16][17][18][19].However, these mammalian muscle cells and tissue-integrated devices require precise environmental control to keep the contractile ability of the muscle cells. The culture medium must be replaced often, and the pH and temperature of the medium must be strictly controlled around 7.4 • C and 37 • C, respectively [20,21]. On the other hand, the tissues of insects are generally robust over a much wider range of living conditions compared with those of mammals. Baryshyan et al. [21] and Akiyama et al. [22] selected the insect tissue and the dorsal vessel (DV) as a bio-actuator. The DV is a central pulsating blood vessel located along the back of an insect, and it acts as a heart. This means that the DV autonomously contracts at a constant frequency without any kind of control or stimuli. The DV tissue was used to demonstrate a micro-pillar actuator [22,23], a walking robot [24], and an atmospheric-operable bio-actuator [20], which worked at room temperature for 90 days without medium replacement [23]. In these studies, this kind of actuator was also capable of working at temperatures from 5 to 40 •...