Introduction Root canal sealing materials play a crucial role in an endodontic procedure by forming a bond between the dentinal walls and the gutta-percha. The current study aims to analyse the dentinal tubule penetration and adhesive pattern, including the push-out bond strength of six commercially available root canal sealers. Methodology Eighty-four mandibular first premolars were split into seven groups (and n = 12), Group 1: Dia-Root, Group 2: One-Fil, Group 3: BioRoot RCS, Group 4: AH Plus, Group 5: CeraSeal, Group 6: iRoot SP, Group 7: GP without sealer (control). Two groups were made, one for dentinal tubule penetration and the other for push-out bond strength; the total sample size was one hundred sixty-eight. Root canal treatment was performed using a method called the crown down technique, and for obturation, the single cone technique was used. A confocal laser scanning microscope (Leica, Microsystem Heidel GmbH, Version 2.00 build 0585, Germany) was used to evaluate dentinal tubule penetration, and Universal Testing Machine was utilised to measure the push-out bond strength (Shimadzu, Japan) using a plunger size of 0.4 mm and speed of 1mm/min. Finally, the adhesive pattern of the sealers was analysed by HIROX digital microscope (KH-7700). Statistical analysis was carried out by a one-way Anova test, Dunnet’s T3 test, and Chi-square test. Results Highest dentinal tubule penetration was noticed with One-Fil (p<0.05), followed by iRoot SP, CeraSeal, AH Plus, Dia-Root also, the most negligible value was recorded for BioRoot RCS. Meanwhile, BioRoot RCS (p<0.05) demonstrated the greater value of mean push-out bond strength, followed by One-fil, iRoot SP, CeraSeal, AH Plus and Dia-Root. Regarding adhesive pattern, most of the samples were classified as type 3 and type 4 which implies greater sealing ability and better adherence to the dentinal wall. However, BioRoot RCS revealed the most type 4 (p<0.05), followed by AH Plus, One-Fil, CeraSeal and Dia-Root. Conclusion The highest dentinal tubule penetration was shown by One-Fil compared to other groups. Meanwhile, BioRoot RCS had greater push-out bond strength and more adhesive pattern than other tested materials.