Background: Tympanometry is an objective test of middle ear function that is routinely used in audiology practice for the identification of middle ear dysfunction. The present study was carried out toinvestigate results of tympanometry using 1000 Hz probe tone in neonates (1-7 days) and infants (6- 8 week). Methodology: A cross-sectional study was carried out. Group I included 65 ears of neonates in the age range of 1 to 7 days and Group II included 70 ears of infants in the age range of 6 to 8 weeks. A calibrated GSI Audio Screener was used for performing transient evoked otoacoustic emissions (TEOAEs) and auditory brainstem response (ABR) screening. Madsen-Otoflex 100 middle ear analyzer was used for recording tympanometry. Single component and multicomponent tympanogram was recorded for 1000 Hz probe tone. Type of tympanogram as well as the values for tympanometric peak pressure, peak admittance at tympanic membrane, tympanometric width and equivalent ear-canal volume were recorded. Unpaired-t test was carried out to find significant difference between the two groups with p <0.05 considered as significant value. Results: In Group I (neonates), single component tympanometry showed that 95% ears presented with single peaked tympanogram, 3.33% had notched and 1.67% had flat tympanogram for 1000 Hz probe tone. Analysis of multicomponent tympanometry, revealed 1B1G tympanogram in 96.67% and 3B1G pattern in 3.33%. In Group II (infants), single component analyses revealed 85% ears presented with single peak and 15% ears with notched tympanograms. When analyzed for multi component tympanometric pattern, 53.33% in infants presented with 1B1G pattern, 45% showed 3B1G pattern and 1.66% ears had 3B3G pattern. The age has a significant effect on admittance at the tympanic membrane and tympanometric width only. Conclusion: The type of tympanogram obtained from healthy middle ear varies depending on age. High frequency probe tone recommended while evaluating neonates and infants lesser than 8 weeks. The data obtained can serve as normative for 0 -1 week neonates and 6-8 week infants. Keywords: Tympanogram; Tympanometric peak pressure; Tympanometric width