The study aimed to investigate the impact of the using local skin cooling on arteriovenous fistula cannulation pain and the comfort levels of hemodialysis patients. The single-blind randomized controlled trial was formed with 50 patients between June 20, 2023, and July 31, 2023, in the hemodialysis units of two state hospitals in Turkey. Patients were randomly assigned to either the experimental group (n = 25) or the control group (n = 25) using the block randomization method. This data were collected through the use of the CoolSense device, the "Patient Information Form," the "Verbal Category Scale," and the "General Comfort Scale." Patients in the experimental group received the local skin cooling for five seconds during arteriovenous fistula cannulation, while the control group followed the standard hospital protocol. The study was registered on Clinical Trials (NCT06144801). The study revealed that the application of the local skin cooling during arteriovenous fistula cannulation significantly reduced pain and increased patients' comfort levels, demonstrating an anesthetic effect (p < 0.001). Furthermore, it was observed that as pain levels decreased, patient comfort levels increased (p < 0.001). It is concluded that the using lokal skin cooling is an effective tool for reducing pain and enhancing comfort during arteriovenous fistula cannulation in hemodialysis patients.