This study evaluated the effects of different final irrigation regimens on the dentin tubule penetration of three different root canal sealers using confocal laser scanning microscopy (CLSM). A total of 160 single-rooted extracted mandibular premolar teeth were divided into five groups (
n
=
32
each) according to the solution used in the final rinse protocol, as follows: 17% ethylenediaminetetraacetic acid (EDTA) (group 1), 17% EDTA and 2% chlorhexidine gluconate (CHX) (group 2), 7% maleic acid (MA) (group 3), 7% MA and 2% CHX (group 4), and 5.25% NaOCl (group 5). Two roots from each group were examined under scanning electron microscopy (SEM) to visualize smear layer removal. Experimental groups were then split randomly into three subgroups (
n
=
10
) and obturated using a cold lateral condensation technique with 0.1% rhodamine B-labelled sealers [either AH Plus (group A), EndoREZ (group E), or Tech BioSealer Endo (group T)] and gutta-percha. Specimens were sectioned and observed by CLSM to evaluate the percentage and maximum depth of sealer penetration at the apical, middle, and coronal levels. Statistical comparison was performed on grouped (apical, middle, and coronal segments) and ungrouped data using two-way ANOVA with Bonferroni post hoc test (
p
<
0.01
). AH Plus showed the greatest tubule penetration while Tech BioSealer Endo showed the least. Resin-based sealers displayed deeper and more consistent penetration. CHX irrigation positively influenced sealer tubule penetration.
The Effect of Different Final Irrigation Regimens on the Dentinal Tubule Penetration of Three Different Root Canal Sealers: A Confocal Laser Scanning Microscopy Study In Vitro