Objectives: To design a device that allows the formation of in situ oral biofilm with similar characteristics to those from the dental plaque, overcoming the limitations of previous devices. Study Design: The Intraoral Device of Overlaid Disk-holding Splints (IDODS) was designed and manufactured. To test its validity, five healthy adult volunteers wore them for two and four days allowing the biofilm to grow without any type of distortion. After each period, the thickness, vitality and structure of the formed biofilm were measured with a Confocal Laser Scanning Microscope (CLSM) in combination with a dual fluorescence solution. All volunteers filled out a Likert-type questionnaire to evaluate the device. Results: Mean bacterial vitality in the 2- and 4-day biofilms was 71% and 63%, respectively. Mean thicknesses were 21 µm and 28 µm, respectively. There was predominance in the open and heterogeneous structure whose complexity was ascending as the biofilm matured. The results obtained from the questionnaire were 2/5 in the influence in aesthetics, 3.4/5 in comfort, and 5/5 in ease of maintaining oral hygiene and withdrawal from the oral cavity . Conclusions: A biofilm with optimum characteristics was obtained by IDODS. Its use is associated with good aesthetic and comfort results and is absent of functional limitations, allowing optimal oral hygiene without altering the structure of the in situ oral biofilm.