Multiple-step beams carrying intermediate lumped masses with/without rotary inertias are widely used in engineering applications, but in the literature for free vibration analysis of such structural systems; Bernoulli-Euler Beam Theory (BEBT) without axial force effect is used. The literature regarding the free vibration analysis of Bernoulli-Euler single-span beams carrying a number of spring-mass systems, Bernoulli-Euler multiple-step and multi-span beams carrying multiple spring-mass systems and multiple point masses are plenty, but that of Timoshenko multiple-step beams carrying intermediate lumped masses and/or rotary inertias with axial force effect is fewer. The purpose of this paper is to utilize Numerical Assembly Technique (NAT) and Differential Transform Method (DTM) to determine the exact natural frequencies and mode shapes of the axial-loaded Timoshenko multiple-step beam carrying a number of intermediate lumped masses and/or rotary inertias. The model allows analyzing the influence of the shear and axial force effects, intermediate lumped masses and rotary inertias on the free vibration analysis of the multiple-step beams by using Timoshenko Beam Theory (TBT). At first, the coefficient matrices for the intermediate lumped mass with rotary inertia, the step change in cross-section, left-end support and right-end support of the multiple-step Timoshenko beam are derived from the analytical solution. After the derivation of the coefficient matrices, NAT is used to establish the overall coefficient matrix for the whole vibrating system. Finally, equating the overall coefficient matrix to zero one determines the natural frequencies of the vibrating system and substituting the corresponding values of integration constants into the related eigenfunctions one determines the associated mode shapes. After the analytical solution, an efficient and easy mathematical technique called DTM is used to solve the differential equations of the motion. The calculated natural frequencies of Timoshenko multiple-step beam carrying intermediate lumped masses and/or rotary inertias for the different values of axial force are given in tables. The first five mode shapes are presented in graphs. The effects of axial force, intermediate lumped masses and rotary inertias on the free vibration analysis of Timoshenko multiple-step beam are investigated.